Tumor derived exosomes as carrier of pro-angiogenic cargo from different cancer models promotes neo-angiogenesis.
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These proteins were initially referred as to hemagglutinins or phytoagglutinins, since they were originally isolated from extracts of plants [1]. The first hemagglutinin isolated by Stillmark was extracted from seeds of the castor tree (
Thirty-one years after Stillmark, James B. Sumner, isolated from jack bean (
In 1907, Landsteiner and Raubitschek analyzed the hemagglutination of red blood cells from different animals by various seeds extracts. They found that the relative hemagglutinating activity was quite different for each extract tested [1]. However, it was only in the 1940s that Willian Boyd and Karl Renkonen, working independently, made the important discovery of human blood groups specificity for hemagglutinins. They found that crude extracts of two leguminous plants,
The specific interaction between lectins and carbohydrates of erythrocytes played a crucial role in the investigations of the antigens associated with the ABO blood group system. In the subsequent decade, Morgan and Watkins found that the agglutination of type A erythrocytes by extracts of
Around thirty years after Boyd, the research on lectins reached the molecular level studies. It was clear the need to a better understanding on the structural aspects of lectins. Then, in 1972 Edelman and colleagues established the primary sequence of ConA [6]. In the same year, Edelman’s group and independently Karl Hardman with Clinton Ainsworth, solved the 3D structure of ConA by X-ray crystallography [7,8].
In 1954 Boyd and Shapleigh proposed the term lectin, from the Latin verb
Lectins were early defined as carbohydrate-binding proteins of nonimmune origin that agglutinate cells or as carbohydrate-binding proteins other than antibodies or enzymes. However, these definitions were updated, since some plant enzymes are fusion proteins composed of a carbohydrate-binding and a catalytic domain, for instance, type 2 RIPs, such as ricin and abrin, are fusion products of a catalytically active A-chain (which has the N-glycosidase activity) and a carbohydrate-binding B-chain, both linked by a disulfide bond [10]. Furthermore, there is in nature carbohydrate-binding proteins possessing only one binding site and, therefore, are not capable of precipitating glycoconjugates or agglutinating cells [11].
Thus, in 1995 Peumans and Van Damme proposed the most suitable definition for lectins. According to the “new” definition, all plant proteins that possess at least one noncatalytic domain that binds reversibly to a specific mono- or oligosaccharide are considered as lectins [12,13].
Lectins are proteins widely distributed in nature such in microorganisms, plants, animals and humans, acting as mediators of a wide range of biological events that involve the crucial step of protein-carbohydrate recognition, such as cell communication, host defense, fertilization, cell development, parasitic infection, tumor metastasis, inflammation, etc [14-15].
Peumans and Van Damme classified the plant lectins according to their overall structure.
The most thoroughly investigated lectins have been isolated from plants, particularly that extracted from members of the Leguminosae family. Legume lectins are a large group of proteins that share a high degree of structural similarity with distinct carbohydrate specificities. The subtribe Diocleinae (Leguminosae) comprises 13 genera, mostly of them from the New World. However, only 3 of these genera (i.e.
Concerning the biological activity, legume lectins are considered as enigmatic proteins. Despite the philogenetic proximity as well the high degree of similarity shared between them, they possess different biological activities such as histamine release from rat peritoneal mast cells, lymphocyte proliferation and interferon-γ production, peritoneal macrophage stimulation and inflammatory reaction as well as induction of paw edema and peritoneal cell immigration in rats [16].
Significant progress has been reached in last years in understanding the crucial roles of lectins in several biological processes [17]. The importance of lectins as biotechnological tools has been established early in the studies involving its biological application. In 1960 a major step in immunology was given in order to determine the role of these proteins on the lymphocytes cell division. It was found that the lectin of the red kidney bean (
In addition to immunological studies, recent works have been investigated the influence of lectins in the field of microbiology, since lectins can be considered as valuable tools to verify the role of interaction between the pathogen and carbohydrates present in host cells and its importance to disease development. For instance, it has been proposed that the pathogen
Currently, malignancies are considered a major problem in the public health, especially given their increasing incidence and prevalence rates observed in recent decades. In this context, the malignant tumors, or cancers, account for approximately 7.8 million deaths per year, thus becoming the second greatest cause of death worldwide, only behind the cardiovascular disease [23].
The cancer can be defined as a set of more than 100 diseases that have in common the uncontrolled growth of cells, which invade tissues and organs and can spread to other body regions [23]. Thus, both the processes of cellular mutation that affects the neoplastic cell and metastasis, involve a series of genetic changes that culminate in modifications in the pattern of several receptors and signaling molecules present on the cell surface [24].
Carbohydrates are biomolecules that have enormous potential for encoding biological information. These combined-molecules (Glycoproteins and Glycolipids) are responsible for different biological interactions between the cell and the extracellular environment [26]. Regarding the neoplastic cells, the glycosylation of these proteins and lipids is changed, which generates membrane signaling molecules capable of inducing several processes directly related to tumor progression such as cell adhesion, angiogenesis, cellular mitosis and metastasis, in addition, in some cases, be responsible for inhibition of apoptosis induction triggered by the cells of the immune system [26].
Certain changes in glycans occur frequently in neoplastic cells and may be considered "tumor-specific", establishing a correlation between the stage of disease progression and prognosis of the same [25]. Some classic examples of these changes are the antigens of the ABH and Lewis system. ABH antigens are not expressed in cells of healthy human colon but significantly expressed on tumor cells [27]. Since the antigen Lewisy can be observed in several carcinomas and has been correlated with poor prognosis in breast tumors [28]. Another example is the glycosylated antigens sialyl-LewisA and sialyl-Lewisx, which are significantly up-regulated in carcinomas of the colon and appear to be related to tumor progression [29].
Thus, due to the intrinsic role of carbohydrates in the tumorigenesis, the glycosylation process as well as the identification of glycosylated antigens have been intensively focused, given the fact that glycosylation of antigens can vary extensively depending on the stage of the disease, which can provide, when properly identified, a better possibility of correct diagnosis and treatment.
Because the peculiar characteristic of specific binding to carbohydrates, lectins have been used as tools to identify aberrant glycans expressed by neoplastic cells. Such methods have been essential to obtain a more precise diagnosis that allows a more accurate prognosis.
Several methods regarding the use of lectins as tools for recognizing aberrant glycans have been proposed in recent days [30,31,32]. The technique most common and widespread is the use of lectins in immunohistochemical assays.
In this context, the study conducted by [30], showed that leguminous lectins from
Another recent methodology was addressed by [33]. This methodology exploits the fact that glycoproteins produced by cancer cells have altered glycan structures, although the proteins themselves are common, ubiquitous, abundant, and familiar. However, as cancer tissue at the early stage probably constitutes less than 1% of the normal tissue in the relevant organ, only 1% of the relevant glycoproteins in the serum should have altered glycan structures [34]. With that in mind, the strategy to approach the detection of these low-level glycoproteins is based in: (a) a quantitative real-time PCR array for glycogenes to predict the glycan structures of secreted glycoproteins; (b) analysis by lectin microarray to select lectins that distinguish cancer-related glycan structures on secreted glycoproteins; and (c) an isotope-coded glycosylation site-specific tagging high-throughput method to identify carrier proteins with the specific lectin epitope [33].
Therefore, further analyses of lectins as biomarkers have been undertaken to improve our understanding of the processes involved in malignant tumor formation. As well as enable us to acquire new methods of identification of neoplastic cells at an early stage, enabling a better prognosis with appropriate treatment and low cost.
Apoptosis is a mechanism by which cells undergo death to control cell proliferation or in response to irreparable DNA damage. It is featured by unique morphological and biochemical changes, such as nucleus condensation and margination, membrane blebbing, and inter-nucleosomal DNA cleavage [35]. As the type I programmed cell death (PCD), apoptosis occurs through two major pathways, the extrinsic pathway triggered by the Fas death receptors, and the mitochondria-dependent pathway that brings about the release of Cytocrome
Several studies have demonstrated that lectins can induce apoptotic cell death. In the mitochondrial-dependent pathway, ConA treatment results in a decrease of mitochondrial membrane potential, and thus collapsing mitochondrial transmembrane potential. Cyto
In [38], it was evaluated the pro-apoptotic activity of a lectin isolated from
An interesting study conducted by [39] showed the pro-apoptotic caspase-dependent activity of the lectin isolated from
Despite the apoptotic activity of several lectins have been demonstrated in different studies, the precise mechanism of how this process is triggered as well the mode of internalization is unknown until the present date.
The inflammation is a nonspecific event of immune response that occurs in reaction to any type of tissue injury. This process is capable of triggering a series of physiological changes such as increased blood flow, elevated cellular metabolism, vasodilation, release of soluble mediators, extravasation of fluids and cellular influx [41].
The continuity of cell recruitment and tissue damage in addition to chemical mediators released by the injured tissue as well as resident cells on site activate various mechanisms, in turn, induce the migration of more immune cells as well as increasing local tissue perfusion [42].
Both, acute and chronic inflammations have specific characteristics and the innate immune system plays a central role, since it mediates the initial response. Infiltration of innate immune cells, specifically neutrophils and macrophages, characterizes the acute inflammation, while infiltration of T lymphocytes and plasma cells are features of chronic inflammation [41,42].
As discussed previously, carbohydrates can act as the intermediates of communication in biological processes such as differentiation, proliferation and certain cell–cell interactions that are crucially important in both physiological and pathological phenomena [43,44]. The information contained in the enormous variety of oligosaccharide structures normally conjugated to proteins or lipids on cell surfaces (glycocodes) is recognized and deciphered by a specialized group of structurally diverse proteins, the lectins [44].
Galectins (formerly “S-type lectins”), an evolutionarily conserved family of endogenous animal lectins, share unique features, including their highly conserved structure, exquisite carbohydrate specificity, and ability to differentially regulate a myriad of biological responses [45].
Although galectins have been implicated in many biological activities, most of the functional studies reported to date link galectins to early developmental processes, such as neovascularization, regulation of immune cell homeostasis and inflammation [44,46,47]. Through deciphering glycan-containing information about host immune cells or microbial structures, galectins can modulate a diversity of signaling events that lead to cellular proliferation, survival, chemotaxis, trafficking, cytokine secretion and cell−cell communication [46,47].
These findings are extremely important because they demonstrate the importance of the glicocodes in the process of cell recognition and inflammation. In this context, plant lectins have been widely used to understand the pro-inflammatory mechanisms, as well as the design of new compounds with pro-healing effect.
An immune system is a system of biological structures and processes within an organism that protects against disease. In order to function properly, an immune system must detect a wide variety of agents, from viruses to cancer cells, and distinguish them from the organism\'s own healthy tissue [41]. The immune system is composed of many cells and molecules that act in a complex and harmonious way with the ultimate goal of annihilating the aggressive factor [42].
In the immune system, two phases of activity can be clearly established: the innate immune response and the adaptive immune response. In the innate immune response, there is the activity of cells and cytokines in a nonspecific way with the main purpose to annihilate quickly the local damaging agents. At this stage, we highlight the neutrophils, eosinophils, basophils and macrophages, cells with well-established activities but with the common function of production and release of cytokines. These cytokines are molecules with various functions in the inflammatory process, such as chemotaxis, activation of certain cell groups and increased tissue perfusion [48].
On the other hand, the adaptive immune response is composed by another set of cells that acts in a more specific way, the lymphocytes. Such cells are responsible for producing antibodies specific for certain invading microorganisms and the activation of mechanisms of apoptosis in abnormal cells [49].
Thus, the use of molecules capable of inducing cell recruitment as well as cytokine production and lymphocytes proliferation is of special scientific interest.
Korean mistletoe (
A recent study [51] demonstrated the immunomodulatory activity of ConBr, a lectin isolated from
Regarding the activity of lectins on lymphocytes, a recent study [52] evaluated the effect of lectin extracted from seeds of
The biological function of carbohydrates in inflammation events is well-defined. In this context, proteins that bind specifically to such glycans are of great interest because of their possible functions and applications in biotechnological studies.
Recently, researches have undertaken efforts at the possible pro-healing activity of some lectins [53,54]. This goal is supported by the fact that such molecules may interfere with the inflammatory process. This effect is not yet fully elucidated, however peculiar and interesting results can be observed.
Experiment conducted in a murine model, employing the lectin isolated from
Although promising, this issue requires further studies to better characterize the mechanisms involved in pro-healing role played by lectins.
To date, there are fewer than 100 publications describing the presence of lectins in marine red, green and brown macroalgae. Moreover, and in marked contrast to higher land plant lectins, marine algal lectins have been isolated and characterized at a much lower pace since the first report of haemagglutinating activity in these organisms appeared more than 46 years ago [56]. Thereafter, other studies describing the presence and/or purification of algal lectins were reported by groups from England [57], Japan [58], Spain [59], United States [60] and Brazil [61].
Currently, the presence of lectins was analyzed at about 800 algae species. However, this number is still small, considering that there are thousands of species of marine algae. Together, the research shows that approximately 60% of the analyzed species show hemagglutinating activity. The number of positive species could be higher since in the first screenings the authors used a limited number of red blood cells and without enzyme treated erythrocytes.
The improvement in the methodologies of both, extraction and hemagglutination assays could increase the number of positive species. In fact, there appears to be coincidence that the rabbit erythrocytes treated with papain are most suited for the hemagglutinating activity detection in marine macroalgae [62,63].
Although marine algal lectins show proteinaceous content similar to lectins from terrestrial plants, they differ in some aspects. Early publications on this issue, reported that in general, lectins from algae have low molecular masses, no affinity for monosaccharides, strong specificity for complex oligosaccharides and/or glycoproteins. Moreover, they appear to have no requirement for metal ions, showing high content of acidic residues and even in high concentrations tend to stay in the monomeric form [64,65,66]. However there are a few reports showing that some of these molecules may be inhibited by simple sugars and are cation dependent as showed for the lectins from the green marine alga genus,
The classical methods used to purify marine algae lectins include methods such as protein precipitation (using salt or ethanol), liquid chromatography (especially affinity) and electrophoresis [69,71]. Ion exchange chromatography has been effectively used in the isolation of lectins from seaweed, mainly in initial stages in purification. In this step, the lectins were separated from pigments present in the extracts [66,72,73]. In the protein extracts, phycobilins are co-extracted with lectins, becoming an undesirable contaminant in the purification process [72].
Lectins from marine algae
To date, only 31 lectins from Rhodophyceae and 17 lectins from Chlorophyceae were isolated and characterized. The virtual absence of lectins isolated from brown algae (Phaeophyta) is mainly due to the amount of polyphenols present in plants. It is known that polyphenols are released in extraction and that these compounds and their oxidation products, quinones, bind tightly to proteins [94] causing a false hemagglutination [83,95].
Even with the increase in the publications related to marine algae lectins, biochemical and structural information on algal lectins is scarce and from only a few species, and hence the functional and phylogenetic classification of these lectins remains unclear. The available structural information indicates the existence of different carbohydrate-binding proteins in the marine algae investigated. Moreover, the complete amino acid sequences of only 14 algal lectins have been determined. In red marine algae,
In same year, [97] reported the primary structure of
On the other hand, the lectins isolated from
Recently [90] reported that the aggregation of cell organelles of
The lectin BPL-2 is a 17 kDa protein specific to D-mannose (ref). The authors found no similarity with others proteins in specific databases. The BPL-3 [92] possesses specificity to
Another observation is that a large number of sequenced algal lectins have the presence of cysteinyl residues or the duplication of internal domain. Still a lot of work needs to be done on the structure of algal lectins, since a few amount of primary structures has been determined. Further structural studies will contribute to understanding the differences in their biochemical characteristics as well as to the evolutionary aspects upon lectin presence in land plants and marine algae.
There are few studies in the literature about the biotechnological applicability of lectins from marine algae. Probably due to low rentability of lectins obtained through the purification processes. It is noteworthy that the majority of lectins isolated so far were extracted from red algae, which are rich in carbohydrates and not in proteins. Moreover, during the extraction of algae proteins, phycobiliproteins are extracted simultaneously, and therefore the addition of steps to remove these phycobiliproteins causes losses of other proteins, among these are the lectins.
However, several studies on biological applications of algae lectins demonstrate that these molecules have an additional benefit; they are molecules with low molecular weight and may be less antigenic when used in biological models.
In cancerology, it was demonstrated that a lectin from the red marine algal
In current studies, Span 80 vesicles (a potential type of nonionic vesicular drug delivery system) with ESA and PEGylated (EPV) lipids immobilized, showed hemagluttinating activity similar to free ESA and decreased the viability of Colo201 cancer cells
The lectins isolated from the red marine algae
Concerning the mitogenic activity, the lectin from the red marine alga
The antibacterial effect was too evaluated. The lectins from the red marine algal
Red marine algae lectins from
Concerning the antiviral effects, a lectin named KAA-2 from the red marine algal
Biofilms are microbial complex communities established in a wide variety of surfaces and are generally associated with an extracellular matrix composed by several types of polymers [121]. This type of microbial association can develop on biotic and abiotic surfaces, including living tissues, medical devices and/or industrial water piping systems and marine environments [122,123,124].
Bacterial infections involving biofilm formation are usually chronic and often present a arduous treatment [125,126,127]. The growth and proliferation of microorganisms inside the biofilm provides reduction or prevents the penetration of various antimicrobial agents [128,129] and thus become extremely difficult or impossible to eradicate them [130,131,132]. For some antibiotics, the concentration required to eliminate the biofilm can be up to a thousand times higher than required to planktonic form of the same specie [127].
The ability of microorganisms to form pathogenic cell aggregates is a worldwide concern. In attempt to remedy this problem, pharmaceutical companies associated to research groups work avidly to the development of new options for the treatment of infections caused by bacteria organized in biofilms.
Biofilm formation is a process in which bacteria has a change in lifestyle, it goes from a state unicellular in suspension to a multicellular sessile, where the growth and cell differentiation results in structured communities. The biofilm begins with the setting free of microorganisms in a given area. The first microorganisms adhere initially by weak interactions, mainly of the van der Waals forces [133]. If the colonies are not immediately removed from the surface, they can anchor by cell adhesion molecules existing in the pili and / or flagella [134]. The first colonies facilitate the arrival of other cells through adhesion sites and begin to build the matrix that will form the biofilm. Only a few species are able to adhere to a surface
According to [136], the biofilm development occurs through three events. At first, there is a distribution of fixed cells in surface through cell motility. Then, occurs the proliferation of fixed cells by division, expanding to upward and sides forming agglomerated of cells, similar to the formation of colonies on agar plates [137]. Finally, clusters of cells attached to the biofilm are recruited by the action of the environment itself to the development of other biofilms, reaching a climax community [136]. These general stages provide guidance to the study of biofilms by bacteria furniture, although many details of regulation of this process may vary between species.
One of the biofilm-forming components of great importance in the maintenance of cell clusters is the matrix polymeric called EPS (
The first step to biofilm formation, the early adhesion, is considered essential for colonization and infection by pathogenic bacteria. Macromolecules surfaces are directly involved in this stage [142]. Proteins known as adhesins are able to recognize specific polysaccharide substrate present on the surface to be colonized, for example, the presence of carbohydrates existing in the film of saliva that covers the teeth in the oral environment [143]. Glycidic epitopes present on surfaces of microorganisms (early colonizers) can also be recognized by these proteins to mediate an event known as coaggregation, which will start the formation of a multi-species community [144].
One etiological factor for the development of teeth biofilms is the adhesion of pathogenic bacteria in the dental enamel [145]. However, the microorganisms are not deposited directly on the tooth surface, but bind to a thin acellular layer composed of salivary proteins and other macromolecules that cover the tooth surfaces called acquired pellicle [146,147].
The acquired pellicle is formed by glycoproteins and carbohydrates that serve as receptors for bacteria containing proteins with glucan-binding domains [148]. Bacteria interact with the film by several specific mechanisms, including the interaction lectin-like involving the bacterial adhesins and receptors existing in the pellicle [148,149]. Next, other bacteria can adhere to the film as well as the bacteria pre-existing in the biofilm [150].
The event coaggregation is a phenomenon widely observed in diverse microbial communities [151,152,153]. Cells can interact in suspension, forming cell aggregates, as well as connect directly on biofilm in the process of formation. In the first case, plancktonic cells recognize specifically species genetically distinct creating the coaggregates. In other situations, coaggregates in the form of secondary colonizers can adhere on biofilm in development, a process known as coadhesion. Both cases have an important role in the integration and establishment of a mature biofilm [154].
Thus, carbohydrate residues have an important role in formation and maintainability of microbial biofilms. They act as mediators of the binding between bacteria and the surface that will serve as substrate for biofilm formation [155], as well as site of interaction between microorganisms to form cell aggregates [156,157]. Furthermore, through EPS matrix, maintains the biofilm attached in the surface, conferring a greater resistance to antimicrobial agents in general [158,159].
Molecules able to bind specifically and selectively to carbohydrates have a key importance in the development of research related to microbial biofilms. Thus, lectins have been shown as powerful tools for analyze the glycidic structures of those aggregates from microbial origin [160,161].
Studies of microbial biofilms through the interaction with lectins have two main objectives: visualization and characterization of polymeric matrix (EPS) formed by different species of microorganisms [162,163] and inhibition of oral biofilm formation by blocking the bacterial binding sites present in the pellicle of saliva in the form of glycoproteins and / or carbohydrates [164].
The application of lectins in the characterization of EPS is already widely exploited by many research groups. Lectin of wheat germ (WGA) was used as a marker of
In 1980s was developed a system called ELLA (
Recent studies demonstrated the characteristics and distribution of glycoconjugates in cyanobacteria biofilm using lectins with different specificities. In this study the authors stated that the distribution of carbohydrates in the matrix is very variable. Based on lectin specificity, glycoconjugates produced by cyanobacteria biofilm contained mainly fucose, N-acetyl-glucosamine or -galactosamine and sialic acid [169].
Lectins may be a suitable antiadhesion agent for
We would like to thank CNPq, CAPES, FUNCAP and UFC for financial support. AHS, BSC, CSN, EHT, KSN are senior investigators of CNPq.
Tumor microenvironment interacts with tumor cells, creating an environment to suppress or contribute towards tumor development and progression [1]. For the tumor development, inflammation and angiogenesis are the processes which play vital roles from initial to the advanced stages of cancer [2]. Extreme angiogenesis and neo-angiogenesis play a fundamental role in tumor progression, which is driven by various pro-and anti-angiogenic factors [3]. There are different ways for tumor cells to communicate with adjacent cells/tissues for facilitating tumor progression; one of these is through exosomes [4, 5]. Exosomes can transport various biomolecules like DNA fragments, mRNAs, noncoding RNAs, proteins, and lipids from a source cell to target/recipient cells that can enhance angiogenesis, which play a significant role in cancer progression [6]. There are evidences that various noncoding RNAs, particularly microRNAs and long non-coding RNAs (lncRNAs) play significant role in the regulation of angiogenesis [7]. Thus, alteration of angiogenesis has become a striking approach for development of effective cancer therapy [1].
Prior to the discovery of exosomes it was assumed that the transmission of information between mammalian cells occurs in an indirect manner. In 1983, two pioneer studies carried out on the differentiation of reticulocytes into mature erythrocytes, reported release of transferrin receptors into extracellular space in form of small vesicles, which were later termed as “exosomes” by R.M. Johnstone [6, 8, 9, 10]. EVs are vesicles enclosed with phospholipid bilayer secreted in the extracellular matrix. Initially, they were initially considered as “garbage dumpsters” but now they are popularly being referred as “signal boxes” [11]. The presence of extracellular vesicles in solid tissue, physiological fluid, and cell culture supernatants has been demonstrated by a number of studies [12]. EV’s are broadly categorized into different subtypes like microsomes, microvesicles, retrovirus-like particles and apoptotic bodies, different from each other on the basis of size, surface markers and their mode of biogenesis [13]. Extracellular vesicle is a collective term for exosomes and microvesicles. Microvesicles originate from through outward budding and fusion of plasma membrane whereas, exosomes are released via endocytosis and fusion with plasma membrane [14]. Exosomes are the smallest (30–100 nm) subpopulation of EVs. CD9, CD63 and Alix are the specific surface markers for these exosomes [13]. Exosome serve as important cell communication regulators and have gained more attention among all the diverse types of extracellular vesicles because they represent a more homogenous set of vesicular population more closely representing the parent cell of origin [15].
Exosomes are endosome derived extracellular vesicles. Multivesicular endosomes (MVEs) or multivesicular bodies (MVBs) are secreted via intracellular secretion pathway, from the plasma membrane. Early endosomes develop into MVBs which fuse with the cell membrane and release the exosomes or else undergoes degradation in lysosomes and autophagosomes. They are cup-or disc-shaped when observed under electron microscopy having a diameter of 30–150 nm [11, 16]. Various proteins and molecules like (ALIX, VPS4, and TSG101) are some of the major proteins involved in exosome biogenesis, content assembly and their secretion via endosomal sorting complex [16]. Exosome biogenesis supposedly occurs via two major pathways: Endosomal sorting complexes required for transport (ESCRT) dependent and ESCRT independent. The ESCRT dependent process includes ESCRT complex (0, I, and II) which are involved in recognizing and sequestering the ubiquitinylated proteins on the endosomal membrane. Exosomes are formed by membrane remodeling, involving bud formation by invagination of this endosomal membrane [17]. ESCRT independent pathway involves tetraspanins such as CD63 and lipid metabolism enzymes like neutral sphingomyelinase (nSMase) and rab family protein consisting of more than 60 GTPases that regulate intracellular trafficking of exosomes [16]. Anchoring of MVBs and transportation of different exosomes is carried out by different RAB subtypes proteins. Early endosome transportation involves RAB5 and RAB21 proteins to mediate endocytosis pathway from early to late endosome and then to lysosome for degradation involves RAB7 protein. Tumor-associated vesicle trafficking requires a vital protein that is RAB27 and it is highly expressed in several tumors. Other than this, various RAB proteins which include RAB 3,11,26,27, 35, 37 and RAB 38 are linked with the exocytic pathway of vesicle trafficking [11]. RAB27 helps in the release of exosomes from mature endosomes enriched in TSG101, ALIX and CD63 whereas RAB11 & RAB35 are associated with the release of early nuclear endosomes which are enriched with PLP, Wnt and TfR. Finally, MVBs fused with the plasma membrane and exosomes are excreted out in the extracellular environment [12]. Diagrammatic representation of exosome biogenesis and secretion has been shown in Figure 1.
Schematic representation of exosome biogenesis and secretion from eukaryotic cells. Exosome’s formation starts with endocytosis, which involves inward budding of plasma membrane, leading to the formation of early and late endosomes. Further, small vesicles are generated by inward budding of late endosomes and forming multivesicular bodies (MVBs). The ultimate fate of MVBs can be either fusion with lysosome for degradation or fusion with plasma membrane to release exosomes. The exosome formation from MVBs proceeds through ESCRT-dependent and ESCRT-independent pathways. ESCRT-dependent pathway involves various ESCRT proteins like (ESCRT 0, I, II, and III) and ESCRT-independent includes lipids (ceramide) and the tetraspanins.
Exosomes are nanovesicles enriched with a repertoire of biomolecules like proteins, nucleic acids and lipids [16]. Exosomes are dynamic and heterogeneous in nature with respect to their content which majorly depends on their cellular origin, pathological and physiological state of the parent cells. Exosomes from different cell types are enriched specifically in proteins like Alix, Tsg101, integrins, Rab GTPases, tetraspanins (CD9) and (CD63), MHC class II proteins and heat shock proteins (HSP90, HSP70), which alsoserve as exosome marker proteins [16, 18]. Besides these, exosomes are also enriched with double-stranded DNA’s and RNA population of different classes such as microRNA (miRNA), long noncoding RNA (lncRNA) [19]. ExoCarta and Vesiclepedia (http://microvesicle.org/), databases have cataloged the RNA, protein and lipid content of exosomes derived from different sources.
Tumor derived exosomes (TEXs) have been shown to play a significant role in tumor progression by accelerating angiogenesis [20]. New blood vessel formation occurred when angiogenic signaling pathways are activated by tumor-derived exosomes, when they are up taken by normal ECs [21]. Exosomal cargo once internalized into recipient cells present in the tumor microenvironment, can regulate their fate, function, and phenotype [22, 23]. Tumor cell derived exosomal cargo can activate/inhibit the various signaling pathway in ECs via receptor-ligand interaction [24]. There are several studies represent multiple avenues in which cancer-derived exosomes exert pro-angiogenic effects on ECs. Till date, the different signaling pathways that are involved in exosomes-induced angiogenesis are poorly known. However, the exosomal cargo which is involved in tumor progression and angiogenesis have been documented. Role of TEXs cargoes which is involved in tumor angiogenesis is showed in Figure 2. Also, a list of all mRNAs, proteins, and noncoding RNAs which are found in TEXs for regulating tumor angiogenesis are listed in Table 1.
Tumor derived exosomes as carrier of pro-angiogenic cargo from different cancer models promote neo-angiogenesis. Tumor-derived exosomes are enriched in proangiogenic proteins, mRNAs, miRNAs, and long noncoding RNAs which are transferred to recipient endothelial cells and activate various angiogenic signaling pathways involved in different angiogenesis process via cell proliferation, migration, and invasion.
Exosomal cargo | Tumor type | Type of study ( | Cell lines | Target cell | Mechanisms | Function | References | |
---|---|---|---|---|---|---|---|---|
EGFRVIII | Glioma cells | Both | U373Viii | U373 and HUVECs | Increase in the VEGF gene expression, by activating the MAPK and Akt pathways | Pro-angiogenesis | [25, 26] | |
Dll4 | Glioma cells | Both | U87MG | HUVEC | Inhibition of notch signaling | Pro-angiogenesis | [27] | |
POU3F3 lncRNA | Glioma cells | A172, U87-MG, U251 and T98G | HBMVECs | Increasing the expression of bFGF, VEGFA and bFGFR in ECs | Pro-angiogenesis | [22] | ||
HOTAIR lncRNA | Glioma cells | A172 | HBMVECs | Increase in the VEGFA expression of ECs | Pro-angiogenesis | [28] | ||
CCAT2 lncRNA | Glioma cells | A172, U87-MG, U251, and T98G | HUVECs | Increase in the expression of VEGFA and other angiogenic signaling molecules of ECs and decrease in the apoptosis process | Pro-angiogenesis | [29] | ||
IL-8, PDGF | Glioblastoma | U87MG | ECs | PI3K/AKT signaling | Pro-angiogenesis | [30] | ||
VEGF-A | Glioblastoma | GSC | Brain microvascular ECs | Enhancement in angiogenic potential of brain ECs | Pro-angiogenesis | [31] | ||
miR-148a-3p | Glioblastoma | U-138-MG, U251-MG, and HEK-293 T | HUVECs | Activating the EGFR/MAPK signaling pathway by inhibiting ERRFI1 | Pro-angiogenesis | [32] | ||
miR-182-5p | Glioblastoma | U-251MG, H4, A-172, U-118MG, LN-18, and U-87MG | HUVECs | Targeting Kruppel-like Factor 2 and 4 | Pro-angiogenesis | [33] | ||
miR-10b | Breast cancer | MCF-7 and MM-231 | HMLE | Suppression of HOXD10 and KLF4 proteins level | Promotes cell invasion | [34] | ||
miR-373 | Breast cancer | MCF-7 and MM-231 | ECs | Wnt/β-catenin signaling | Pro-tumorigenesis | [35] | ||
miR-122 | Breast cancer | Both | MCF-10A and MM-231 | Normal cells in pre metastasic niche | Downregulation of PKM | Promotes metastasis, before angiogenesis | [36] | |
miR-497 | Breast cancer | Both | MCF-7 | HUVECs | Decrease in the expression of VEGF and HIF-1 | Anti-angiogenesis | [37] | |
AnxA2 | Breast cancer | Both | MCF10A and MM-231 | Macrophages and ECs | Generation of plasmin | Pro-angiogenesis | [38] | |
miR-210 | Breast cancer | Both | 4 T1 | ECs | Upregulation of VEGF | Pro-angiogenesis | [39] | |
miR-145 | Breast cancer | Both | MDA-MB-231 | HUVECs | STIM1 promotes angiogenesis by reducing exosomal miR-145 which targets IRS1 | Pro-angiogenesis | [40] | |
NA | Breast cancer | MCF-7 and MM-231 | ADSCs | SMAD pathway | Pro-angiogenesis | [41] | ||
miR-135b | Multiple myeloma | Both | RPMI8226, KMS-11 and U266 | ECs | Suppression of FIH-1 | Pro-angiogenesis | [42] | |
Angiogenin, bFGF, VEGF | Multiple myeloma | Both | ST33MMVT and RPMI8226 | ECs, bone marrow stromal cells | Activation of P53, N-terminal kinase, C-jun and STAT3, | Pro-angiogenesis | [43] | |
miR-9 | Melanoma | SK23 | ECs | JAK-STAT pathway | Pro-angiogenesis | [44] | ||
IL-6, VEGF, and MMP-2 | Melanoma | HTB63, Mewo, and A375 | ECs | WNT5A signaling pathway | Pro-angiogenesis | [45] | ||
GM-CSF, HIF-1α, HIF-2α | Melanoma | NA | ECs and M1/M2 macrophages | Upregulation of VEGF expression | Pro-angiogenesis | [46] | ||
Tetraspanin Tspan8 (D6.1A) | Pancreatic cancer | Both | BSp73AS | ECs | Upregulation in the expression of MMP, VEGF, and VEGFR | Pro-angiogenesis | [47, 48] | |
Wnt4 | Colorectal cancer | Both | HT29 and HCT116 | ECs | Wnt/β-catenin pathway | Pro-angiogenesis | [49] | |
lncRNA UCA1 | Pancreatic cancer | PANC-1, MIA PaCa-2, BxPC-3, Aspc-1, Sw1990, and HEK293T | HUVECs | AMOTL2/ERK1/2 Signaling Pathway | Pro-angiogenesis | [50] | ||
M-phase-related transcripts | Colorectal cancer | SW480 | ECs | Modulation of M-phase of cell cycle and activation of cell proliferation | Initiate angiogenesis | [51] | ||
miR-21 | Lung cancer | SV40 | HUVECs | Upregulation of VEGF | Pro-angiogenesis | [52] | ||
miR-23a | Lung cancer | NCI-H1437, H1648, H1792 and H2087 | HUVECs | Exosomal miR-23a increased angiogenesis and vascular permeability by targeting prolyl hydroxylase and tight junction protein ZO-1 | Pro-angiogenesis | [53] | ||
miR-141 | Small cell lung cancer | H446 and H1048 | HUVECs | Exosomal miR-141/KLF12 pathway | Pro-angiogenesis | [54] | ||
Profilin 2 | Small cell lung cancer | H446 | HUVECs | t PFN2 activated Smad2/3 in H446 and pERK in ECs | Pro-angiogenesis | [55] | ||
Vasorin | Hepatocellular carcinoma | HepG2 | HUVEC | Promote cell proliferation and migration | Pro-angiogenesis | [56] | ||
Angiopoietin-2 | Hepatocellular carcinoma | Both | Hep3B, SNU182, SNU387, Li7 and MHCC97H | HUVECs | Tie2-independent pathway | Pro-angiogenesis | [57] | |
miR-1290 | Hepatocellular Carcinoma | Hep3 B and HepG2 | HUVECs | miR-1290-Induced proangiogenic phenotype via targeting SMEK1 | Pro-angiogenesis | [58] | ||
NA | Renal cancer | 786-0 | HUVEC | Upregulation of VEGF, expression and downregulation of hepaCAM | Pro-angiogenesis | [59] | ||
NA | Renal cancer | 786-0 | 786-0 | Increase in the expression of CXCR4 and MMP-9 | Enhance migration and invasion | [60] | ||
CA9 | Renal cancer | 786-0 | HUVEC | Increasing the MMP-2 expression | Pro-angiogenesis | [61] | ||
miR-549a | Renal cancer | Both | 786-0 and 293T | HUVECs | Exosomal miR-549a affects angiogenesis and endothelial cell migration by silencing HIF1α in HUVECs | Pro-angiogenesis | [62] | |
miR-27a | Renal clear cell carcinoma | 786-0, RPTEC and HEK293T | HUVECs | RCCC-derived miR-27a-loaded exosomes inhibit SFRP1 expression and accelerate tumor angiogenesis in RCCC | Pro-angiogenesis | [63] | ||
EDIL-3 | Bladder cancer | TCC-SUP, T24, and SV-HUC | HUVEC | Promote cell proliferation and migration | Pro-angiogenesis | [24] | ||
miR-181a | Papillary thyroid cancer (PTC) | Both | BCPAP and K1 | HUVECs | Hypoxic PTC-secreted exosomes delivered miR-181a that inhibits DACT2 via downregulating MLL3, leading to YAP-VEGF-mediated angiogenesis | Pro-angiogenesis | [64] | |
miR-21 | Head and neck squamous cell carcinoma | Both | FaDu | CD14+ human monocytes | Increasing the expression of M2 polarization markers of TAMs | Pro-angiogenesis | [65] | |
ICAM-1, CD44v5 | Nasopharyngeal carcinoma | C666-1, NP69 and NP460 | HUVEC | Src kinase, ERK1/2 kinase, p38 MAPK, RhoA/ROCK, and eNOS | Pro-angiogenesis | [66] | ||
PFKFB-3 | Nasopharyngeal carcinoma | CNE2 | HUVEC | Increasing in the production of Fru-2,6-P2 and lactate | Pro-angiogenesis | [67] | ||
HMGB3 | Nasopharyngeal carcinoma | Both | CNE1, CNE2, 5-8 F, 6-10B and NP69 | HUVECs | HMGB3-containing nEXOs accelerated angiogenesis in vitro and in vivo | Pro-angiogenesis | [68] | |
FAM225A lncRNA | Esophageal squamous cell carcinoma cells | ECA109, TE-1, KYSE150, and KYSE-410, and HET-1A | HUVECs | Sponging miR-206 thus derepressing its targets NETO2 and FOXP1 thereby activating PI3K/Akt/NF-κB/Snail axis | Pro-angiogenesis | [69] | ||
miR-130a | Gastric cancer | Both | SGC-7901 | HUVEC | Downregulation of c-MYB | Pro-angiogenesis | [70] | |
NA | Chronic myeloid leukemia | Both | K562 | HUVEC | Src pathway | Pro-angiogenesis | [71] | |
IL-8 | Chronic myeloid leukemia | Both | LAMA84 | HUVEC | MAPK signaling | Pro-angiogenesis | [72] | |
miR-92a | Chronic myeloid leukemia | K562 | ECs | Targeting integrin-α5 | Pro-angiogenesis | [73] | ||
miR-210 | Chronic myeloid leukemia | K562 | ECs | Downregulation of EFNA3 | Pro-angiogenesis | [74] | ||
miR-21 | Chronic myeloid leukemia | Both | K562 LAMA84 | HUVEC | Downregulation of RhoB | Anti-angiogenesis | [75] | |
TGF-β | Prostate cancer | LNCAP, DU145, and PC3 | Fibroblasts | SMAD-dependent signaling | Pro-angiogenesis and pro-tumorigenesis | [76] | ||
C-Src, IGF-IR, FAK | Prostate cancer | DU145, PC3 and C4-2B | ECs | Upregulation of VEGF | Pro-angiogenesis | [77] | ||
VEGF | Ovarian cancer | CABAI | HUVEC | Acts through its tyrosine kinase receptors | Pro-angiogenesis | [78] | ||
CD147 | Ovarian cancer | CABAI, A2780, OVCAR3 and SKOV3 | HUVEC | Upregulation of MMP and VEGF | Pro-angiogenesis | [79] | ||
ATF2, MTA1, SARS, ROCK1/2 | Ovarian cancer | CAOV3 | HUVEC | Upregulation of VEGF and HIF-1α | Pro-angiogenesis | [80] | ||
miR-221-3p | Cervical cancer | CasKi, SiHa, HeLa and SW756 | MVECs | CC cells-derived exosomes harboring miR-221-3p enhanced MVEC angiogenesis in CC by decreasing MAPK10 | Pro-angiogenesis | [81] | ||
miR-141-3p | Ovarian cancer | SKOV-3a | HUVECs | Activating the JAK/STAT3 and NF-κB signaling pathways | Pro-angiogenesis | [82] | ||
PTCH 1, SMO, SHH, Ihh | Cervical cancer | SiHa, HeLa and C33a | HUVECs | CC cells-derived exosomes promote pro-angiogenic response in endothelial cells via upregulation of Hh-GLI signaling and modulate downstream angiogenesis-related target genes | Pro-angiogenesis | [83, 84] | ||
TIE2 | Cervical cancer | SiHa, HeLa and THP1 | HUVECs | TIE2-high tumor cells deliver TIE2 to macrophages to induce TIE2-expressing macrophages via exosomes | Pro-angiogenesis | [85] | ||
RAMP2-AS1 lncRNA | Chondrosarcoma cells | SW1353 | HUVECs | Sponging miR-2355-5p thus derepressing its target VEGFR2 thereby increasing angiogenic cell surface receptors | Pro-angiogenesis | [86] | ||
miR-92a-3p | Retinoblastoma | Both | WERI-Rb1 | HUVECs | Exosomally delivered miR-92a-3p modulates angiogenesis by targeting KLF2 | Pro-angiogenesis | [87] | |
miR-155 | Burkitt’s lymphoma | Raji | ARPE-19 | Upregulation of VEGF-A expression via VHL/HIF-1α pathway | Pro-angiogenesis | [88] |
Tumor derived exosomes as carrier of pro-angiogenic cargo from different cancer models promotes neo-angiogenesis.
Exosomes derived from glioblastoma cells are known to carry different mRNAs, miRNAs and angiogenic factors which interacts with ECs and thus stimulate angiogenesis. Kucharzewska et al. demonstrated export of pro-angiogenic factors IL-8 and PDGF through exosomes derived from the hypoxic glioma cells and thus induce endothelial proliferation and cell migration by activating the PI3K/AKT signaling pathway [30]. Exosomes from glioblastoma cells showed enrichment of different non-coding RNAs that include, microRNAs (miRNAs): miR-148a-3p, miR-182-5p; long non-coding RNAs (lncRNAs): POU3F3, HOTAIR, CCAT2 in the regulation of glioma cell angiogenesis [22, 28, 29, 32, 33]. Exosomes derived from glioma cells are also known to carry pro-angiogenic proteins such as EGFRvIII, VEGF-A and DII4 which are important for tumor growth, survival and angiogenesis through the activation of Akt and MAPK signaling pathways [25, 26, 27, 31].
Breast cancer derived-exosomes transfer majorly pro-angiogenic microRNAs: miR-10b, miR-101, miR-105, miR-122, miR-145, miR-210 and miR-373 responsible for tumor invasion, metastasis and lead to angiogenesis [34, 35, 36, 39, 40, 41]. However, Wu et al. found that exosomes secreted from breast cancer cells loaded with miR-497 are responsible for anti-angiogenesis by downregulating the VEGF and HIF-1 [37]. Maji et al. have observed that Annexin A2 was transferred via breast cancer exosomes to ECs and induces the process of vascularization and angiogenesis through the tissue plasminogen activator (tPA)-dependent manner
Multiple myeloid cancer cells derived exosomes are known to carry miR-135b and responsible for tube formation in ECs by suppressing its target FIH-1 [42]. Wang et al. observed that various pro-angiogenic factors are released into the exosomes derived from multiple myeloma cells such as angiogenin, bFGF and VEGF that promote tumor growth [43].
In a study conducted by Zhuang et al. demonstrated that exogenous miR-9 can advance tumor angiogenesis by downregulating the SOCS-5 levels, which can discordantly regulate the JAK-STAT signaling pathway [44]. Hood et al. have observed exosomes released from melanoma cells stimulate the expression of HIF-1α, HIF-2α and GM-CSF, which leads to angiogenesis in endothelial cells [46]. Moreover, Ekstrom et al. showed that the WNT5A signaling promotes the exosomal secretion from melanoma cells containing immunomodulatory and pro-angiogenic factors such as IL-6, MMP-2 and VEGF [45].
Pancreatic adenocarcinoma produced exosomes having high levels of tetraspanin Tspan8 (D6.1A) that promote migration, proliferation and sprouting in ECs. Moreover, these exosomes also help in maturation of endothelial progenitor cells [47, 48]. Guo et al. showed that lncRNA UCA1 was exported through exosomes derived from the hypoxic pancreatic cancer cells are responsible for angiogenesis via miR-96-5p/AMOTL2 signaling pathway [50].
Studying the exosomes from the colorectal carcinoma demonstrated that these exosomes carry pro-angiogenic factors Wnt 4, which helps in angiogenesis of ECs through Wnt/β-catenin pathway [49]. Hong et al. found that the exosomes released from SW480 colorectal cancer cell lines are loaded with M-phase related transcripts such as RAD21, CDK8, and ERH and regulate M-phase of the cell cycle and promotes proliferation and in turn enhance angiogenesis [51].
Exosomes derived from small cell lung cancer (SCLC) cells are found to be enriched with miR-21 and miR-23a, which is correlated with the pro-angiogenic activities in ECs [52, 53]. A study of Mao et al. demonstrated that exosomes from SCLC cells are responsible for pro-angiogenic effect via miR-141/KLF12 pathway in targeted ECs [54]. In another recent study, Profilin2 protein was transferred from the lung cancer cells via exosomes and leads to angiogenesis by activating the t-PFN2 dependent pERK pathway in endothelial cells [55].
Vasorin (VASN), a type I transmembrane protein has an effective role in tumor progression and angiogenesis, was secreted by exosomes of hepatocellular carcinoma cells (HCC) and promotes the migration of HUVEC cells [56]. In another study of Xie et al. showed that angiopoietin-2 protein is transferred to ECs from HCC cells via exosomes and responsible for pro-angiogenesis [57]. Recently, it was found that miR-1290 is also released from the HCC cells through exosomes and responsible for angiogenesis by inducing the miR-1290 induced pro-angiogenic phenotype in endothelial cells, by targeting the SMEK1 [58].
Zhang et al. demonstrated that exosomes derived from renal cancer cell enhances angiogenesis by upregulating the expression of VEGF and downregulating the hepaCAM expression in ECs [59]. Moreover, exosomes derived from renal cancer 786-0 cells promotes invasion and migration of the endothelial cells through upregulation of chemokine receptors CXCR4 and MMP-9 [60]. A recent study of Hou et al. observed that the exosomes derived from renal clear cell carcinoma (RCCC) are loaded with miR-27a and inhibits SFRP1 expression which leads to accelerated angiogenesis in HUVECs [63].
Beckham et al. observed that the exosomes derived from urine of patients with bladder cancer and high-grade bladder cancer cell lines contain an angiogenic factor. Epidermal growth factor (EGF)-like repeats and discoidin I-like domain-3 (EDIL-3) that facilitate cell proliferation and migration which leads to angiogenesis in endothelial cells. EDIL-3 activated EGFR signaling overrule this EDIL-3 induced bladder cell migration [24].
In a recent study by Wang et al. observed that miR-181a is delivered by hypoxic PTC-secreted exosomes inhibits DACT2 by downregulating MLL3, leading to YAP-VEGF-mediated angiogenesis by increasing proliferation and forming capillary-like network in HUVECs. Further, angiogenic potential of hypoxic PTC-secreted exosomes was confirmed in-vivo, which was reversed in presence of hypoxic miR-181 inhibitor [64].
Chan et al. showed that nasopharyngeal carcinoma (NPC) derived exosomes are supplemented with pro-angiogenic factors, ICAM-1 and CD44v5, which helps in angiogenesis of endothelial cells [66]. In another study by Gu et al. recognized a vital role of PFKFB-3 in NPC derived exosomes, which helps in migration, proliferation and angiogenesis of HUVECs [67]. Exosomes derived from FaDu cells are highly enriched with miR-21, captured by monocytes present in the TME and responsible for increasing the expression of M2 polarization of TAMs markers, which helps in tumor progression by regulating the tumor invasiveness and angiogenesis [65]. In a recent study, it was observed that a nuclear protein HMGB3 is transferred to endothelial cells via exosomes released from NPC cells and responsible for accelerated angiogenesis
Zhang et al. demonstrated that exosomes released from esophageal squamous cells are enriched with lncRNA FAM225A, which accelerates esophageal squamous cell carcinoma progression and angiogenesis by sponging miR-206. Further, they showed the upregulation of NETO2 and FOXP1 expression when FAM225A absorbed the miR-206 thereby activating PI3K/Akt/NF-κB/Snail axis [69].
Exosomes derived from gastric cancer cell are enriched with miR-130a and plays a central role in tumor angiogenesis. They showed that exosomal miR-130a is able to facilitate angiogenesis by downregulating the c-MYB, which is an important transcription factor in different biological processes [70]. In another study by Li et al. demonstrated that exosomes released from irradiated gastric cancer cells promote invasiveness and proliferation of endothelial cells [89].
LAMA84 a human CML cell line releases exosomes and are able to trigger diverse signaling pathways in ECs, leading to enhanced expression of important angiogenic factor IL-8 [72]. Umezu et al. observed that exosomes from leukemia cells can transport miR-92a into ECs and responsible for enhanced tube formation and migration by downregulation of integrin-α5 [73]. In another study, it was found that leukemia cell derived exosomes are able to induce tube formation in HUVECs by activating Src [71]. It has been observed that exosomes released from K562 leukemia cells are loaded with miR-210 downregulate the receptor tyrosine kinase ligand, Ephrin A3 (EFNA3) [74]. However, in contrast, Taverna et al. showed that curcumin treatment deeply changes the molecular properties of exosomes released by leukemia cells, in particular, deplete the exosomes of the pro-angiogenic proteins and leads to enrichment of proteins with anti-angiogenic activity and miR-21 [75].
Exosomes derived from prostate cancer cells are known to carry TGF-β1 protein, which can induce the differentiation of recipient fibroblasts to myofibroblasts [76]. In a study by DeRita et al., showed that prostate cancer cell exosomes were loaded with, IGF-IR, FAK and c-src, which could promote tumor angiogenesis [77].
Taraboletti et al. demonstrated that exosomes from ovarian cancer cells are known to carry pro-angiogenic growth factor VEGF, which helps in interaction between tumor and endothelial cells and is very important for angiogenesis [78]. Ovarian cancer exosomes are enriched with pro-angiogenic protein CD147, ATF 2, MTA1, SARS and ROCK1/2. They observed that these proteins can enhance the expression of vital angiogenic factors like VEGF, HIF-1α and MMPs and resulting in the enhanced angiogenesis of HUVECs [79, 80]. Additionally, Masoumi-Dehghi et al. observed that exosomes from ovarian cancer cells are enriched in miR141-3p, which helps in angiogenesis by activating the JAK/STAT and NF-kB signaling pathways [82].
Cheng et al. demonstrated that microarray analysis revealed that exosomes released from chondrosarcoma cells carried lncRNA RAMP2-AS1, which promotes HUVECs migration, proliferation, and tube formation which leads to angiogenesis through miR-2355-5p/VEGFR2 axis, thereby regulating the angiogenic ability of endothelial cells. Successive experiments showed that RAMP2-AS1 knockdown could decrease the pro-angiogenic effect of exosomes released from chondrosarcoma cells [86].
Recently a study conducted by Chen et al. demonstrated that exosomes released by human retinoblastoma cell line WERI-Rb1, were enriched inmiR-92a-3p. The study, predicted that Krüppel-like factor 2 (KLF2) might activate target of miR-92a-3p, using bioinformatics tools & analysis. Thus, exosomal miR-92a-3p was found to modulate tumor angiogenesis by targeting KLF2 [87].
A study performed by Yoon et al. observed that miR-155 is transported from EBV-positive Burkitt’s lymphoma cells derived exosomes which could induces angiogenesis in retinal epithelial pigment (RPE) cells (ARPE-19) by upregulation of transcriptional and translational levels of VEGF A via VHL/HIF-1α pathway. Thus, study demonstrated that miR-155 accumulation through exosomes affect nearby recipient cells [88].
Zhang et al. observed that exosomes released from cervical cancer cells harboring miR-221-3p, which accelerate the MVEC migration, proliferation, invasion and angiogenesis in cervical cancer cells by regulating MAPK10 [81]. In another study performed by Bhat et al. showed that cervical cancer exosomes were highly enriched with upstream proteins of hedgehog-GLI signaling includes, PTCH1, SMO, SHH and Ihh [83]. Also, they observed that these cervical cancer exosomes facilitate pro-angiogenic endothelial reconditioning through transfer of Hedgehog-GLI signaling components [84].
The discovery of exosomes as natural carriers of different mRNAs, miRNAs and lncRNAs makes them a suitable candidate as therapeutic drug vehicles and drug carriers to target cancer cells and modulation of tumor microenvironment. Recent advance in the field reveals several success stories (Table 2). The manipulation of exosomes as drug carriers provides significant advantage for example their nonimmunogenic nature [95]. Exosomes are also known to carry different cell surface molecules due to which they have a commendable ability to transgress numerous biological barriers, such as the BBB (blood-brain barrier). They are highly stable in blood, which permits them to perform long distance intercellular communication [96]. Clinical data from various studies revealed that progression of cancer can be delayed or prevented when tumor angiogenesis is blocked [97]. So, angiogenesis during tumor development has now become the major emphasis of study and angiogenesis inhibition is evolving as a new method to treat cancer [98]. Recent investigations reported that exosomes can decrease or increase angiogenesis based on their molecular content. Thus, there is a lot of promise in developing engineered exosomes to transport numerous biological and synthetic genetic materials that can modify the expression of various genes involved in tumor angiogenesis [99]. For example, Ohno et al. demonstrated that modified exosomes carrying EGF or GE11 on their surface can deliver miR let-7a (tumor suppressor miR) to EGFR expressing breast cancer cells in RAG2−/− mice model. Their previous investigation showed that GE11-exosomes which delivered miR-let 7a, effectively downregulated HMGA2 expression in cancer cells [90]. This study verifies that exosomes can be used as drug delivery vehicle to transport their cargo efficiently to the target cells. Exosomes have capability to act as carriers for delivering different small interfering RNAs (siRNAs) for targeted cancer treatment. Exosomes having HGF siRNA packed inside them can be transported into gastric cancer cells, where they downregulate the HGF expression [91]. Liu et al. demonstrated that exosomes are able to transport antisense RNA targeted to miR-150, which induces the expression of VEGF. They established that the neutralization of miR-150 downregulates the VEGF levels in mice and blocked angiogenesis [92]. Gupta et al. have shown that the bone marrow stromal cells (BMSCs) are involved in the tumor progression by secreting different pro-angiogenic factors, bFGF and VEGF [100]. In another study, it was observed that the miR content of exosomes derived from old and young BMSCs was different from each other. Young BMSC exosomes were highly enriched with miR-340, which inhibited the angiogenesis through HGF/c-MET signaling pathway in ECs. The antiangiogenic effect of older BMSCs was remarkably enhanced, when miR-340 was transferred to older BMSC exosomes that was highly expressed in young BMSC exosomes. Therefore, this investigation indicates the exosome-based cancer therapy via replenishment of miRNAs of exosomes [94]. The Arg-Gly-Asp (RGD) sequence containing peptide specifically bounds to αVβ3 integrin and plays an important role in endothelial cell survival, migration and angiogenic growth. In a study performed by Wang et al. showed successful binding of the RGD sequence containing peptide to the exosomal membrane surface and thereby binding of the αVβ3 integrin on the surface of angiogenic blood vessel. Thus, engineered exosomes are emerging as a new probable therapeutic motor for angiogenesis therapy [99]. In another study, it has been observed that curcumin treated CML cells released the exosomes, which are highly enriched with miR-21, which is further transferred to ECs and downregulates the expression of RhoB [75]. Docosahexaenoic acid (DHA) is a polyunsaturated omega-3 fatty acid (PUFA) and popularly known for its anti-cancer and anti-angiogenesis properties. A group of researchers demonstrated that exosomes released from the DHA-treated breast cancer cell lines are highly enriched with miRs, including miR-21, miR-27a/b, miR-23b, miR-320b, let-7 and let-7a, which are well known for their anti-angiogenic properties. They observed the increased expression of these miRs when exosomes were co-incubated with the endothelial cells. Collectively, the exosomes show a strong therapeutic potential as natural nano carrier [93].
Exosomal cargos | Study models | Study Outcome | References |
---|---|---|---|
let-7a miR | Breast cancer | Secreted exosomes delivered miR-let7a to the breast cancer cells expressing EGFR and inhibited cancer growth by blocking angiogenesis | [90] |
HGF siRNA | Gastric cancer | Exosomes decrease the tumor growth and angiogenesis in gastric cancer by delivering hepatocyte growth factor siRNA (HGF siRNA) | [91] |
Antisense RNA targeted to miR-150 | NA | Downregulated the expression levels of VEGF in mice and blocked angiogenesis | [92] |
miR-21, miR-23b, miR-27a/b, miR-320b, let-7 and let-7a | Breast cancer | DHA treated exosomes have altered miRNA content that have anti-angiogenic properties in breast cancer | [93] |
miR-340 | Old Bone Marrow Stromal Cells (BMSCs) | Exosomes having miR-340, inhibits angiogenesis through HGF/c-MET signaling pathway in ECs | [94] |
miR-21 | Chronic Myeloid Leukemia (CML) | Exosomes transferred miR-21 to ECs and downregulated the expression of RhoB | [75] |
Engineered exosomes as anti-angiogenic drug carriers in different cancer models.
Herein, we have emphasized the current advances in the roles of tumor derived exosomes in cancers of different origins in tumor angiogenesis. Exosomes could modulate the angiogenic programming in target cells by transferring the angiogenic cargoes that include different mRNAs, miRNAs, lncRNAs and proteins. Angiogenesis is a very complex process in which aberrant growth of tumor and its metastasis occurs. So, the inhibition of angiogenesis is a pivotal point to control the progression of cancer. In spite of increasing amount of information about tumor derived exosomal cargo and changes prompted by them on target cells, the complexity of exosomal cargoes remains to be fully elucidated. There are several limitations and road blockers in the significance of exosomes in cancer therapy. These specifically pertain to exosomal yield, exosomes efficacy and specificity of targeting for effective cancer therapy. This field is yet elusive to assess the effect of exosomes on tumor angiogenesis and use them as potential means for different cancer therapies. So, future investigations should focus on identifying the fundamental exosomal cargoes and the mechanisms behind differential loading of different bioactive molecules, whose role could be implemented for designing non-invasive procedures to detect exosomes for cancer diagnosis and prognosis as well as development of effective therapeutic approaches based on exosomes.
Not Applicable.
The authors declare that there are no competing/conflicts of interest.
Financial support from Science and Engineering Research Board Department of Science and Technology, Government of India (DST-SERB (EMR/2017/004018/BBM)) and Institution of Eminence University of Delhi (Ref. No./IoE/2021/12/FRP) to ACB and grant from CCRH to ACB:SC:KT (17-51/2016–2017/CCRH/Tech/Coll./DU-Cervical Cancer.4850) and Indian Council of Medical Research (ICMR-ICRC (No.5/13/4/ACB/ICRC/2020/NCD-III), are thankfully acknowledged. Study was partly supported by Junior Research Fellowship to TT (764/(CSIR-UGC NET JUNE 2019) and Senior Research Fellowship to AC [573(CSIR-UGC NET JUNE 2017)] by University Grants Commission (UGC), Senior Research Fellowship to NA (09/045(1622)/2019-EMR-I) and JY (09/045(1629)/2019-EMR-I) by Council of Scientific and Industrial Research (CSIR); Junior Research Fellowship to DJ (09/0045/(11635)/2021-EMR-1) and AC (09/0045(12901)/2022-EMR-1).
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Although their expected number is more than 2.2–3.8 million, only 120,000 taxa have been identified so far. Basidiomycetes are very large group of fungi including mushrooms, toad stools, puff balls, earth stars, polypores, and rust and smut fungi. Previously, these fungi were identified only by morphological characters that have been considered as variable due to environmental factors. Literature shows that many fungi are misidentified due to phenotypic changes. Molecular methods including phylogenetics prove to be successful aids along with traditional methods for correct identification of these fungi and these have revolutionized fungal reclassification. Many fungal taxa have been shifted to other groups of fungi after their phylogenetic analysis. So, many DNA markers can be used to solve such problems.",book:{id:"6880",slug:"recent-advances-in-phylogenetics",title:"Recent Advances in Phylogenetics",fullTitle:"Recent Advances in Phylogenetics"},signatures:"Samina Sarwar, Qudsia Firdous and Abdul Nasir Khalid",authors:[{id:"249556",title:"Dr.",name:"Samina",middleName:null,surname:"Sarwar",slug:"samina-sarwar",fullName:"Samina Sarwar"},{id:"262216",title:"Ms.",name:"Qudsia",middleName:null,surname:"Firdous",slug:"qudsia-firdous",fullName:"Qudsia Firdous"},{id:"262218",title:"Prof.",name:"Abdul Nasir",middleName:null,surname:"Khalid",slug:"abdul-nasir-khalid",fullName:"Abdul Nasir Khalid"}]},{id:"63795",title:"Phylogenetics",slug:"phylogenetics",totalDownloads:2204,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"Describing the diversity of living beings has always instigated man. The classification proposed by Aristotle today seems naïve and unnatural, but it lasted from ancient Greece until the publication of the Linnaeus Systema Naturae in 1758. Although quite accurate, the taxonomic classification proposed by naturalist Carl Linnaeus did not consider the evolutionary relationships between living beings. This view, although prior to Charles Darwin, only gained deserved prominence after On the Origin of Species. Only in the twentieth century, a new area founded by Hennig, phylogenetic systematics was implemented, and with this, a series of useful methods in the construction of phylogenetic trees arose, as maximum parsimony, neighbor joining, UPGMA, maximum likelihood, and Bayesian inference. With the advancement of information technology, phylogenetic analyses have become more sophisticated and faster. The algorithms used in the analysis programs have become more complex and realistic, favoring the addition of substitution models. The application of these data and the greater facility in generating nucleotide and amino acid sequences allowed the comparison previously unimaginable, for example, between bacteria and eukaryotes. In this way, the history of the advances of phylogenetic knowledge is confused with the greater knowledge about the origin of life.",book:{id:"6880",slug:"recent-advances-in-phylogenetics",title:"Recent Advances in Phylogenetics",fullTitle:"Recent Advances in Phylogenetics"},signatures:"Eliane Barbosa Evanovich dos Santos",authors:[{id:"250217",title:"M.Sc.",name:"Eliane",middleName:null,surname:"Evanovich",slug:"eliane-evanovich",fullName:"Eliane Evanovich"}]},{id:"39794",title:"Missense Mutations in GDF-5 Signaling: Molecular Mechanisms Behind Skeletal Malformation",slug:"missense-mutations-in-gdf-5-signaling-molecular-mechanisms-behind-skeletal-malformation",totalDownloads:2583,totalCrossrefCites:0,totalDimensionsCites:8,abstract:null,book:{id:"2535",slug:"mutations-in-human-genetic-disease",title:"Mutations in Human Genetic Disease",fullTitle:"Mutations in Human Genetic Disease"},signatures:"Tina V. Hellmann, Joachim Nickel and Thomas D. 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He received his Ph.D. in Molecular Biology with his thesis “Genetic variability of the tick-borne encephalitis virus in natural foci of Novosibirsk city and its suburbs.” His primary field is molecular virology with research emphasis on vector-borne viruses, especially tick-borne encephalitis virus, Kemerovo virus and Omsk hemorrhagic fever virus, rabies virus, molecular genetics, biology, and epidemiology of virus pathogens.",institutionString:"Russian Academy of Sciences",institution:{name:"Russian Academy of Sciences",country:{name:"Russia"}}},{id:"310962",title:"Dr.",name:"Amlan",middleName:"Kumar",surname:"Patra",slug:"amlan-patra",fullName:"Amlan Patra",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/310962/images/system/310962.jpg",biography:"Amlan K. Patra, FRSB, obtained a Ph.D. in Animal Nutrition from Indian Veterinary Research Institute, India, in 2002. He is currently an associate professor at West Bengal University of Animal and Fishery Sciences. He has more than twenty years of research and teaching experience. He held previous positions at the American Institute for Goat Research, The Ohio State University, Columbus, USA, and Free University of Berlin, Germany. His research focuses on animal nutrition, particularly ruminants and poultry nutrition, gastrointestinal electrophysiology, meta-analysis and modeling in nutrition, and livestock–environment interaction. He has authored around 175 articles in journals, book chapters, and proceedings. Dr. Patra serves on the editorial boards of several reputed journals.",institutionString:null,institution:{name:"West Bengal University of Animal and Fishery Sciences",country:{name:"India"}}},{id:"53998",title:"Prof.",name:"László",middleName:null,surname:"Babinszky",slug:"laszlo-babinszky",fullName:"László Babinszky",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/53998/images/system/53998.png",biography:"László Babinszky is Professor Emeritus, Department of Animal Nutrition Physiology, University of Debrecen, Hungary. He has also worked in the Department of Animal Nutrition, University of Wageningen, Netherlands; the Institute for Livestock Feeding and Nutrition (IVVO), Lelystad, Netherlands; the Agricultural University of Vienna (BOKU); the Institute for Animal Breeding and Nutrition, Austria; and the Oscar Kellner Research Institute for Animal Nutrition, Rostock, Germany. In 1992, Dr. Babinszky obtained a Ph.D. in Animal Nutrition from the University of Wageningen. His main research areas are swine and poultry nutrition. He has authored more than 300 publications (papers, book chapters) and edited four books and fourteen international conference proceedings.",institutionString:"University of Debrecen",institution:{name:"University of Debrecen",country:{name:"Hungary"}}},{id:"201830",title:"Dr.",name:"Fernando",middleName:"Sanchez",surname:"Davila",slug:"fernando-davila",fullName:"Fernando Davila",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/201830/images/5017_n.jpg",biography:"I am a professor at UANL since 1988. My research lines are the development of reproductive techniques in small ruminants. We also conducted research on sexual and social behavior in males.\nI am Mexican and study my professional career as an engineer in agriculture and animal science at UANL. Then take a masters degree in science in Germany (Animal breeding). Take a doctorate in animal science at the UANL.",institutionString:null,institution:{name:"Universidad Autónoma de Nuevo León",country:{name:"Mexico"}}},{id:"309250",title:"Dr.",name:"Miguel",middleName:null,surname:"Quaresma",slug:"miguel-quaresma",fullName:"Miguel Quaresma",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/309250/images/9059_n.jpg",biography:"Miguel Nuno Pinheiro Quaresma was born on May 26, 1974 in Dili, Timor Island. He is married with two children: a boy and a girl, and he is a resident in Vila Real, Portugal. He graduated in Veterinary Medicine in August 1998 and obtained his Ph.D. degree in Veterinary Sciences -Clinical Area in February 2015, both from the University of Trás-os-Montes e Alto Douro. He is currently enrolled in the Alternative Residency of the European College of Animal Reproduction. He works as a Senior Clinician at the Veterinary Teaching Hospital of UTAD (HVUTAD) with a role in clinical activity in the area of livestock and equine species as well as to support teaching and research in related areas. He teaches as an Invited Professor in Reproduction Medicine I and II of the Master\\'s in Veterinary Medicine degree at UTAD. Currently, he holds the position of Chairman of the Portuguese Buiatrics Association. He is a member of the Consultive Group on Production Animals of the OMV. He has 19 publications in indexed international journals (ISIS), as well as over 60 publications and oral presentations in both Portuguese and international journals and congresses.",institutionString:"University of Trás-os-Montes and Alto Douro",institution:{name:"University of Trás-os-Montes and Alto Douro",country:{name:"Portugal"}}},{id:"38652",title:"Prof.",name:"Rita",middleName:null,surname:"Payan-Carreira",slug:"rita-payan-carreira",fullName:"Rita Payan-Carreira",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRiFPQA0/Profile_Picture_1614601496313",biography:"Rita Payan Carreira earned her Veterinary Degree from the Faculty of Veterinary Medicine in Lisbon, Portugal, in 1985. She obtained her Ph.D. in Veterinary Sciences from the University of Trás-os-Montes e Alto Douro, Portugal. After almost 32 years of teaching at the University of Trás-os-Montes and Alto Douro, she recently moved to the University of Évora, Department of Veterinary Medicine, where she teaches in the field of Animal Reproduction and Clinics. Her primary research areas include the molecular markers of the endometrial cycle and the embryo–maternal interaction, including oxidative stress and the reproductive physiology and disorders of sexual development, besides the molecular determinants of male and female fertility. She often supervises students preparing their master's or doctoral theses. She is also a frequent referee for various journals.",institutionString:null,institution:{name:"University of Évora",country:{name:"Portugal"}}},{id:"283019",title:"Dr.",name:"Oudessa",middleName:null,surname:"Kerro Dego",slug:"oudessa-kerro-dego",fullName:"Oudessa Kerro Dego",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/283019/images/system/283019.png",biography:"Dr. Kerro Dego is a veterinary microbiologist with training in veterinary medicine, microbiology, and anatomic pathology. Dr. Kerro Dego is an assistant professor of dairy health in the department of animal science, the University of Tennessee, Institute of Agriculture, Knoxville, Tennessee. He received his D.V.M. (1997), M.S. (2002), and Ph.D. (2008) degrees in Veterinary Medicine, Animal Pathology and Veterinary Microbiology from College of Veterinary Medicine, Addis Ababa University, Ethiopia; College of Veterinary Medicine, Utrecht University, the Netherlands and Western College of Veterinary Medicine, University of Saskatchewan, Canada respectively. He did his Postdoctoral training in microbial pathogenesis (2009 - 2015) in the Department of Animal Science, the University of Tennessee, Institute of Agriculture, Knoxville, Tennessee. Dr. Kerro Dego’s research focuses on the prevention and control of infectious diseases of farm animals, particularly mastitis, improving dairy food safety, and mitigation of antimicrobial resistance. Dr. Kerro Dego has extensive experience in studying the pathogenesis of bacterial infections, identification of virulence factors, and vaccine development and efficacy testing against major bacterial mastitis pathogens. Dr. Kerro Dego conducted numerous controlled experimental and field vaccine efficacy studies, vaccination, and evaluation of immunological responses in several species of animals, including rodents (mice) and large animals (bovine and ovine).",institutionString:"University of Tennessee at Knoxville",institution:{name:"University of Tennessee at Knoxville",country:{name:"United States of America"}}},{id:"251314",title:"Dr.",name:"Juan Carlos",middleName:null,surname:"Gardón",slug:"juan-carlos-gardon",fullName:"Juan Carlos Gardón",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/251314/images/system/251314.jpeg",biography:"Juan Carlos Gardón Poggi received University degree from the Faculty of Agrarian Science in Argentina, in 1983. Also he received Masters Degree and PhD from Córdoba University, Spain. He is currently a Professor at the Catholic University of Valencia San Vicente Mártir, at the Department of Medicine and Animal Surgery. He teaches diverse courses in the field of Animal Reproduction and he is the Director of the Veterinary Farm. He also participates in academic postgraduate activities at the Veterinary Faculty of Murcia University, Spain. His research areas include animal physiology, physiology and biotechnology of reproduction either in males or females, the study of gametes under in vitro conditions and the use of ultrasound as a complement to physiological studies and development of applied biotechnologies. Routinely, he supervises students preparing their doctoral, master thesis or final degree projects.",institutionString:"Catholic University of Valencia San Vicente Mártir, Spain",institution:null},{id:"125292",title:"Dr.",name:"Katy",middleName:null,surname:"Satué Ambrojo",slug:"katy-satue-ambrojo",fullName:"Katy Satué Ambrojo",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/125292/images/system/125292.jpeg",biography:"Katy Satué Ambrojo received her Veterinary Medicine degree, Master degree in Equine Technology and doctorate in Veterinary Medicine from the Faculty of Veterinary, CEU-Cardenal Herrera University in Valencia, Spain. She is a Full Professor at the Department of Medicine and Animal Surgery at the same University. She developed her research activity in the field of Endocrinology, Hematology, Biochemistry and Immunology of horses. She is a scientific reviewer of several international journals : American Journal of Obstetrics and Gynecology, Comparative Clinical Pathology, Veterinary Clinical Pathology, Journal of Equine Veterinary Science, Reproduction in Domestic Animals, Research Veterinary Science, Brazilian Journal of Medical and Biological Research, Livestock Production Science and Theriogenology. Since 2014, she has been the Head of the Clinical Analysis Laboratory of the Hospital Clínico Veterinario from the Faculty of Veterinary, CEU-Cardenal Herrera University.",institutionString:"CEU-Cardenal Herrera University",institution:{name:"CEU Cardinal Herrera University",country:{name:"Spain"}}},{id:"309529",title:"Dr.",name:"Albert",middleName:null,surname:"Rizvanov",slug:"albert-rizvanov",fullName:"Albert Rizvanov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/309529/images/9189_n.jpg",biography:'Albert A. Rizvanov is a Professor and Director of the Center for Precision and Regenerative Medicine at the Institute of Fundamental Medicine and Biology, Kazan Federal University (KFU), Russia. He is the Head of the Center of Excellence “Regenerative Medicine” and Vice-Director of Strategic Academic Unit \\"Translational 7P Medicine\\". Albert completed his Ph.D. at the University of Nevada, Reno, USA and Dr.Sci. at KFU. He is a corresponding member of the Tatarstan Academy of Sciences, Russian Federation. Albert is an author of more than 300 peer-reviewed journal articles and 22 patents. He has supervised 11 Ph.D. and 2 Dr.Sci. dissertations. Albert is the Head of the Dissertation Committee on Biochemistry, Microbiology, and Genetics at KFU.\nORCID https://orcid.org/0000-0002-9427-5739\nWebsite https://kpfu.ru/Albert.Rizvanov?p_lang=2',institutionString:"Kazan Federal University",institution:{name:"Kazan Federal University",country:{name:"Russia"}}},{id:"210551",title:"Dr.",name:"Arbab",middleName:null,surname:"Sikandar",slug:"arbab-sikandar",fullName:"Arbab Sikandar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/210551/images/system/210551.jpg",biography:"Dr. Arbab Sikandar, PhD, M. Phil, DVM was born on April 05, 1981. He is currently working at the College of Veterinary & Animal Sciences as an Assistant Professor. He previously worked as a lecturer at the same University. \nHe is a Member/Secretory of Ethics committee (No. CVAS-9377 dated 18-04-18), Member of the QEC committee CVAS, Jhang (Regr/Gen/69/873, dated 26-10-2017), Member, Board of studies of Department of Basic Sciences (No. CVAS. 2851 Dated. 12-04-13, and No. CVAS, 9024 dated 20/11/17), Member of Academic Committee, CVAS, Jhang (No. CVAS/2004, Dated, 25-08-12), Member of the technical committee (No. CVAS/ 4085, dated 20,03, 2010 till 2016).\n\nDr. Arbab Sikandar contributed in five days hands-on-training on Histopathology at the Department of Pathology, UVAS from 12-16 June 2017. He received a Certificate of appreciation for contributions for Popularization of Science and Technology in the Society on 17-11-15. He was the resource person in the lecture series- ‘scientific writing’ at the Department of Anatomy and Histology, UVAS, Lahore on 29th October 2015. He won a full fellowship as a principal candidate for the year 2015 in the field of Agriculture, EICA, Egypt with ref. to the Notification No. 12(11) ACS/Egypt/2014 from 10 July 2015 to 25th September 2015.; he received a grant of Rs. 55000/- as research incentives from Director, Advanced Studies and Research, UVAS, Lahore upon publications of research papers in IF Journals (DR/215, dated 19-5-2014.. He obtained his PhD by winning a HEC Pakistan indigenous Scholarship, ‘Ph.D. fellowship for 5000 scholars – Phase II’ (2av1-147), 17-6/HEC/HRD/IS-II/12, November 15, 2012. \n\nDr. Sikandar is a member of numerous societies: Registered Veterinary Medical Practitioner (life member) and Registered Veterinary Medical Faculty of Pakistan Veterinary Medical Council. The Registration code of PVMC is RVMP/4298 and RVMF/ 0102.; Life member of the University of Veterinary and Animal Sciences, Lahore, Alumni Association with S# 664, dated: 6-4-12. ; Member 'Vets Care Organization Pakistan” with Reference No. VCO-605-149, dated 05-04-06. :Member 'Vet Crescent” (Society of Animal Health and Production), UVAS, Lahore.",institutionString:"University of Veterinary & Animal Science",institution:{name:"University of Veterinary and Animal Sciences",country:{name:"Pakistan"}}},{id:"311663",title:"Dr.",name:"Prasanna",middleName:null,surname:"Pal",slug:"prasanna-pal",fullName:"Prasanna Pal",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/311663/images/13261_n.jpg",biography:null,institutionString:null,institution:{name:"National Dairy Research Institute",country:{name:"India"}}},{id:"202192",title:"Dr.",name:"Catrin",middleName:null,surname:"Rutland",slug:"catrin-rutland",fullName:"Catrin Rutland",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/202192/images/system/202192.png",biography:"Catrin Rutland is an Associate Professor of Anatomy and Developmental Genetics at the University of Nottingham, UK. She obtained a BSc from the University of Derby, England, a master’s degree from Technische Universität München, Germany, and a Ph.D. from the University of Nottingham. She undertook a post-doctoral research fellowship in the School of Medicine before accepting tenure in Veterinary Medicine and Science. Dr. Rutland also obtained an MMedSci (Medical Education) and a Postgraduate Certificate in Higher Education (PGCHE). She is the author of more than sixty peer-reviewed journal articles, twelve books/book chapters, and more than 100 research abstracts in cardiovascular biology and oncology. She is a board member of the European Association of Veterinary Anatomists, Fellow of the Anatomical Society, and Senior Fellow of the Higher Education Academy. 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