Summarization of dysregulated miRNA in ALS.
Abstract
Progressing muscle wasting and dramatic neurodegeneration of upper and lower motor neurons are the initial symptoms of amyotrophic lateral sclerosis (ALS) that eventually cause aetiology or death in quick succession. The functional mechanism of ALS is non-cell autonomous but it strongly influences on non-neural cells including microglia, astrocyte muscles and T cell. In ALS, neurodegeneration is triggered by at least four gene mutations that are not related to any classical signalling pathways, molecular mechanism or known cellular ingredients. MicroRNA is endogenous tiny non-coding RNA, which is required for fine-tuning or micromanaging protein expression post-transcriptionally. In this review, we identified numerous microRNAs and their possible targets in ALS-related genes. These microRNAs misprocess ALS-related protein-coding genes via microRNA-gene circuits. This result sheds a strong link between microRNA and ALS genes. The mechanistic insight of multiple microRNAs related to ALS is required to treat neuro-inflammation and neuro-degradation. It is proposed that the micro-regulation of multiple microRNAs is involved in generation of unique neuroprotective agent against ALS. Therefore, a classical and novel microRNA-mediated therapy might unravel an alternative strategy for ALS-related neurodegeneration. This strategy indeed implicates real promises to illustrate a unique impact for ALS cure.
Keywords
- amyotrophic lateral sclerosis (ALS)
- small microRNA
- hotspot
- microRNA-mediated therapy
- neurodegeneration
1. Overview and introduction
Amyotrophic lateral sclerosis (ALS) is an important neuromuscular disease [1]. ALS was recognized back in 1850 by the English neurophysiologist Augustus Waller for the appearance of shrivelled nerve fibres. In 1869, in a scientific literature it was described and named ALS by the French neurologist Jean-Martin Charcot [2]. ALS, otherwise known as Lou Gehrig’s disease, caused a baseball player, Lou Gehrig, to retire from his peak season. This New York Yankees player was called the iron horse for his contribution in the field (adapted from http://www.hopkinsmedicine.org/neurology_neurosurgery/centers_clinics/als/conditions/als_amyotrophic_lateral_sclerosis.html, http://www.biography.com/people/lou-gehrig-9308266).
Gradual degeneration of motor neurons in the brain and spinal cord is sought after to characterize this disease [1]. After the disease contracts, the motor neurons can no longer deliver impulses to the muscles, resulting in atrophy of muscles and muscle weakness (Figure 1). ALS does not disrupt a person’s intellectual reasoning, vision, hearing or sense of taste, smell and touch. Mostly, ALS does not affect a person’s bowel, sexual or bladder functions. ALS is often referred to as a neurodegenerative syndrome because the disease becomes evident in various patterns. ALS occurs scarcely and spontaneously. Currently, there is no straightforward and classical cure for ALS (adapted from http://www.hopkinsmedicine.org/neurology_neurosurgery/centers_clinics/als/conditions/als_amyotrophic_lateral_sclerosis.html, http://emedicine.medscape.com/article/1170097-clinical).
MicroRNAs are the tiny endogenous non-coding RNA that represses protein synthesis post-transcriptionally by coupling at the 3′ untranslated leader sequences of target mRNAs [4]. MicroRNA micro-manages a broad range of biological process including developmental decisions, cellular differentiation, programmed cell death for pattern formation, and many pivotal roles in different human diseases [5]. At least 500–1000 microRNAs are found in vertebrate [6]. However, each microRNA can target numerous mRNAs [7]. It is suggested that 30–40% of human transcriptome is under control of different miRNA-gene circuits.
Biogenesis, processing and functional mechanism of microRNA are unique. Four different functional processes are involved in microRNA-mediated gene expression where (1) co-translational protein synthesis is disrupted, (2) translation elongation is inhibited, (3) translation product is terminated prematurely and finally (4) translation initiation is disrupted. MicroRNA inhibits their functional targets by sequestering to target mRNAs [8].
Basically, microRNA can be used as a fine-tuner for different gene regulatory networks. Especially numerous microRNAs are expressed in different sites of the brains [9]. They participate in the functional mechanism of brain development. They have important roles in brain morphogenesis, neuronal differentiation, dendrite and spine developments, synaptic structure formation and neuronal plasticity, etc. [10]. Multiple clusters of microRNA play important parts in different acute and chronic pathological disorders in the brain. In this review we have mentioned a set of microRNA that produce a distinct cluster in different ALS-related genes. The dysregulation of these microRNAs collectively misregulates multiple mRNAs related to ALS and other neurological diseases. Multiple microRNAs can simultaneously control several endogenous mRNAs because 3′ UTR of an mRNA can have complementary sites for a cluster of microRNAs; conversely a single microRNA can regulate multiple mRNAs [11]. In protein homeostasis, different cellular events participate with different microRNAs. However, different microRNAs may involve various mediators to control gene expression [12].
2. Symptoms of ALS
“ALS is like a lit candle: it melts your nerves and leaves your body a pile of wax. Often, it begins with the legs and works its way up. You lose control of your thigh muscles, so that you cannot support yourself standing. You lose control of your trunk muscles, so that you cannot sit up straight. By the end, if you are still alive, you are breathing through a tube in a hole in your throat, while your soul, perfectly awake, is imprisoned inside a limp husk, perhaps able to blink, or cluck a tongue, like something from a science fiction movie, the man frozen inside his own flesh. This takes no more than 5 years from the day you contract the disease”. Courtesy from “Tuesdays with Morrie” by Mitch Albom.
Each individual patient may experience symptoms differently. Symptoms may include:
Twitching and cramping of muscles, especially those in the hands and feet
Loss of motor control in the hands and arms
Impaired use of the arms and legs
Weakness and fatigue
Tripping and falling
Dropping things
Uncontrollable periods of laughing or crying
Slurred or thick speech and difficulty in projecting the voice
As the disease progresses, symptoms may comprise:
Shortness of breath
Difficulty in breathing
Difficulty in swallowing
Paralysis (adapted from http://www.hopkinsmedicine.org/neurology_neurosurgery/centers_clinics/als/conditions/als_amyotrophic_lateral_sclerosis.html)
3. Symptom statistics
ALS occurs between the ages of 40 years and 70 years, but the disease can occur at a younger age also. It affects throughout the world without any ethnic, racial or socioeconomic boundaries. ALS is responsible for almost five of every 100,000 deaths in people aged 20 or older. The frequent age for ALS is after 60 years age. The incidence of ALS is five times higher than Huntington’s disease and almost equal to multiple sclerosis. Fifty percent of affected patients live at least three or more years after diagnosis, 20 percent live 5 years or more and up to 10 percent will survive more than 10 years (adapted from http://www.hopkinsmedicine.org/neurology_neurosurgery/centers_clinics/als/conditions/als_amyotrophic_lateral_sclerosis.html).
4. Medical classification of ALS
Making a proper diagnosis in ALS is complicated because symptoms can vary in each patient. Based on the symptom, ALS can be classified in five broad ranges based on the disease symbol:
Classical ALS – characterized by the deterioration of upper and lower motor neurons [nerve cells]. This symptom generally affects more than two-thirds of patients with the disease.
Primary lateral sclerosis [PLS] – in which case the upper motor neurons deteriorate. If the lower motor neurons are not affected within 2 years, the disease usually remains a pure upper motor neuron disease. It is the rarest form of ALS.
Progressive bulbar palsy [PBP] – this condition starts with difficulties in speaking, chewing and swallowing due to lower motor neuron [nerve cell] deterioration. It affects about 25% of those with ALS.
Progressive muscular atrophy [PMA] – in which the lower motor neurons deteriorate. If the upper motor neurons are unaffected within 2 years of contracting the disease, the disease usually remains a pure lower motor neuron disease.
Familial – that affects more than one member of the same family (adapted from http://www.hopkinsmedicine.org/neurology_neurosurgery/centers_clinics/als/conditions/als_amyotrophic_lateral_sclerosis.html).
5. Tiny small RNA or microRNA
It seems the world of non-coding RNAs is expanding like the universe, with the progress of science. After the discovery of miRNA in 1993, our knowledge about miRNA is increasing exponentially.
All miRNAs undergo 5′ capping and 3′ polyadenylation, which are common events in case of mRNA processing also [16]. Intronic miRNAs are produced under host gene promoter [15]. Intergenic miRNAs have own promoter [15]. In the nucleus, genes encoding miRNA are generally transcribed by RNA polymerase II (Pol II) into large primary miRNA transcripts (pri-miRNA) (sized >1 kb) [15–17]. The pri-miRNA is maximally 2.2 kb long [15, 17]. Processing of pri-miRNA:pri-miRNA > pre-miRNA > miRNA [15]. miRNAs bind to the 3′ UTR region of target mRNAs [8]. Once the miRNA binds to a completely complementary region of target mRNA, mRNA gets degraded [18]. miRNA-mediated regulation does not require to have a perfect match with its target-binding region [19]. Only seven-base sequence between second and eighth nucleotides from the 5′ end is called “seed region (sequence)” and a complete match of these sequence is required for degradation of target mRNA[s] [20]. It is believed that the strength of the inhibition varies depending on the sequences [20]. A single miRNA may directly affect the expression of hundreds of proteins at one time and several miRNAs can also target the same mRNA and result in enhanced translational inhibition. It causes post-transcriptional gene control [8].
6. Link between miRNA and ALS
In last few decades, neuroscience has made a remarkable progress. This has led to accumulation of humongous information including neuronal signalling and neuronal circuits. Neuroscience itself is complex and RNA interference (RNAi) molecules have made it much more intricate. The finer details are being searched to cope up with the neurodegenerative disorders. In the postgenomic era, we are still searching mysteries related to non-coding RNA. These small RNA molecules are the controller of our genome.
The microRNA molecules are acting as nodes and links of gene expression. These experts work in various biological functions. Using complementarity against cognate mRNA, this 21–22 nt stretch plays havoc. A miRNA can target several mRNAs, and in combination with other regulatory molecules, it can alter a cell’s expression and activity. Neurology includes networking between the cells of nervous system and other cells of the organism. Direct and indirect targets of one miRNA make a cell’s system much more complex. ALS is a motor neuron disorder and it has been proved in various reports that ALS can be caused due to dysregulation of miRNAs and thus misexpression of proteins in the cells (Figure 2). This chapter deals with the miRNAs which are culprits behind ALS [21].
7. MicroRNA and its biogenesis pathway
miRNAs, after being discovered in 1993 [13], has come a long way and established itself as one of key regulators at the post-transcriptional level. It gets expressed in wide varieties of organism from plants to humans. Many miRNAs are conserved across several species. These regulatory molecules are of 21–25 nt in length. Many components of miRNA-processing machinery can be found in Archaea and Eukarya, which supports its ancient presence [3]. Databases of different miRNAs are increasing day by day because of its association with several biological processes and diseases. miRNAs account for >3% of all human genes [22, 23]. At first it gained its initial importance because of its involvement in developmental biology. Now it has been noticed that normal miRNA population can be dysregulated in different diseases, and thus, they are being used as biomarker to detect diseases. Sequence-specific regulation by miRNA makes it more specific but it can diversify its function by binding to 3′ UTR sites of several mRNAs.
The miRNA genes can be operated from their own promoter or promoter of another gene [15]. Primary miRNA transcripts are transcribed by RNA pol II in nucleus [16, 17]. Primary transcripts are of multiple hairpin loops. In nucleus Drosha along with DGCR8 cleaves at 3′ end to form pre-miRNAs. Pre-miRNAs have a hanging 3′-OH end [24]. 70 nt structure pre-miRNAs get transported to cytoplasm from nucleus by Ran-GTP and Exportin-5 [24]. In the cytoplasm, RNase-III Dicer enzyme cut at the loop structure of the pre-miRNA to produce passenger strand and guide strand [24]. The thermodynamically unstable strand, i.e. guide strand, works as a mature miRNA [25]. These mature miRNAs get loaded into RISC complex [RNA-induced silencing complex]. RISC complex contains several proteins which are needed for ribonucleotide binding and cleaving the target mRNA. Argonaute protein, GW182 are some of the factors of RISC [8]. Once RISC factor is assembled, the miRNA guides the complex to its target on the basis of complementary base pairing. The complementarity is generally restricted to the second to eighth base from 5′ end of miRNA, which is called “seed” region [25]. This seed region binds to 3′ UTR of target mRNA [19, 20]. The fate of target mRNA depends upon the extent of base pairing to the cognate miRNA [19, 20]. The miRNA will direct the destruction of the target mRNA, who has the perfect or near-perfect pairing [19, 20]. If there is multiple partially complementary site, the accumulation of protein from target mRNAs will be inhibited.
8. miRNA in mammals
miRNAs can inhibit translation in many ways like co-translational protein degradation, inhibition of translation elongation, premature termination of translation, inhibition of translation initiation, deposition of mRNAs in cytoplasmic P-bodies and premature ribosome drop-off [8].
9. Post-initiation mechanisms
Studies in the worm
10. Inhibition of translation initiation
There was an observation by a group, where it was observed that the central domain of Argonaute proteins has sequence similarities to the cytoplasmic cap-binding protein
11. miRNA-mediated mRNA decay
Argonaute proteins prevent the formation of the closed-loop mRNA configuration by a murky mechanism that includes de-adenylation. MicroRNAs trigger de-adenylation and de-capping of the target mRNA. Proteins required for this process are components of the major de-adenylase complex [CAF1, CCR4 and the NOT complex], the de-capping enzyme DCP2 and several de-capping activators. Thus, signal for translation is lost.
P-bodies or cytoplasmic processing bodies are nothing but ribosome-depleted areas inside a cell. Recent studies support the evidence that the mRNAs silenced by miRNAs are localized to P-bodies for storage or degradation. Induced by miRISC and RNA helicase activity, remodelled mRNPs may modify the translation initiation complex at the 5′ end of target mRNAs, which may cause translation repression and localization of mRNAs in P-bodies. P-bodies may facilitate the access of the de-capping factors to the cap structure, thus facilitating mRNA degradation. However, with the help of appropriate signals, stored mRNAs residing in P-bodies could be released and returned to the translational machinery of the cell [8, 28].
12. miRNAs in different parts of the human brain
One of the important processes is microRNA-mediated gene regulation, which is fine-tuning the expression of genes essential for the pathway. Using bioinformatics approach, one can easily understand that the cells of human brain are finely tuned by miRNAs. Prediction of miRNA target sites using bioinformatics approach can give us a whole bunch of gene names. But a major weakness of in silico analysis is the lack of experimental validation. For circumventing such problem, a combinative approach can be implemented where target sites are predicted using multiple programmes. Out of miRNAs, which get expressed in the midbrain, cerebellum, hippocampus and frontal cortex, majority of them are from midbrain while almost equal proportions of microRNAs are expressed in the hippocampus and frontal cortex. The microRNAs in the cerebellum are less in number compared to miRNAs expressed in other regions considered. The target sites of the microRNA expressed in different parts of the brain had been predicted on the 35 genes related to ALS initially. The result obtained from miRanda [version 1.0] indicates that 477 target sites were predicted for mRNAs expressed in midbrain as compared to the target sites [411] predicted for miRNAs expressed in the hippocampus (Figure 3). However, total target sites predicted for miRNAs expressed in the cerebellum and frontal cortexes were 175 and 395, respectively (Figure 3). The number of targets for miRNAs expressed in cerebellum is considerably less comparing to binding sites for miRNAs expressed in other areas of the brain. Surprisingly, microRNA targets were distributed in the different regions of the genes and are not limited to only in 3′ UTR. Total 1456 target sites were predicted using miRanda for miRNAs considered in the study. Target sites in the 3′ untranslated region are comparatively less. A significant number of targets were also found in the 5′ UTR of the genes. Earlier, there was occurrence of miRNA target sites in coding region and 5′ UTR of genes was considered as exception in animals. But numerous recent evidences have established that microRNAs can target different regions of a gene and microRNA-based regulation is not confined only to 3′ UTR [9, 29]. Experiments aimed at identification of specific parameter or factor for effective targeting of 3′ UTR by miRNAs have failed considerably. Conversely, the existence of miRNA target sites in ORF of Nanog,
13. Causative factors in ALS
Most of the cases are sporadic ALS [sALS]. Only 10–15% are usually inherited as an autosomal dominant trait and defined as familial [fALS]. They show comparable etiopathology and they affect same neuronal population [31]. Several ALS genes (
A wealth of evidence supports that ALS is a multifactorial and multisystemic disease, characterized by overlapping different mechanisms, transduction pathways and multicellular crosstalk. Transgenic animals expressing human mutant SOD1 (mSOD1) have helped to reveal that perishing of motor neurons is not the only reason behind ALS but there is active contribution of non-neuronal cells like microglia, astrocytes and muscle and T cells, which differently participate to the different phases of the disease [36, 37]. Expression of mSOD1 within the most vulnerable motor neurons primarily causes for disease onset; synthesis of the mutant protein by interneurons also positively participates to disease initiation [38, 39]. Speeding of disease progression is caused by moSOD-1-mediated injury and neighbouring astrocytes and microglia. Schwann [40] and muscle cells play as direct partners of the injured motor axons, either working as recipients or initiators of the initial damage. These characteristics thus properly earned to ALS the definition of non-cell-autonomous disease [41]. Chimeric mice formed by expressing both mutated and non-mutated SOD1 proved that wild-type nonneuronal cells prolong the survival of motor neurons carrying mSOD1 protein. Neighbouring astrocytes, microglia, oligodendrocytes and Schwann cells can cause the degeneration of motor neurons [42, 43]. Compelling evidence indicates that primary mSOD1-expressing astrocytes from mouse, rat and humans cause motor neurons death by releasing toxic factors into the media while sparing interneurons [44–46]. Astrocytes deriving from neuronal progenitor cells, gathered from fALS and sALS cases, exhibit features of non-cell-autonomous toxicity [47]. There are very few studies of non-cell-autonomous toxicity. Astrocytes expressing mutant TDP43 [TDP43-M337V] participate in ALS pathology only through cell-autonomous processes. It has been studied that conditioned medium from astrocytes that express SOD1-G93A, or SOD1-G86R, or TDP43-A315T leads to extensive motor neuron death via non-cell-autonomous mechanisms. Still, there is a long way left to travel and to unravel mystifying stories behind ALS [21] (Figures 1 and 3).
14. The miRNAs involved in ALS regulation
14.1. Effects of miRNA biogenesis related but mutated factors in ALS
Several factors are there, which can affect CNS homeostasis and thereby facilitate neurodegenerative and neuroinflammatory diseases. Perturbation in intricate miRNA network can disrupt neuronal function in cell-autonomous or non-cell-autonomous manner [48–50]. It has been shown that there is cell-type-specific deletion of miRNA-processing enzyme, Dicer in cerebellar Purkinje, or striatal, retinal, spinal and cortical neurons, which leads to degeneration and ataxia [51–54]. Neurodegeneration also occurs after targeted deletion of Dicer in astrocytes, oligodendrocytes and Schwann cells, perhaps because the glial cells have some effects on neuronal survival [55–58] (Table 1). Experimental results show that deletion of Dicer directly in spinal motor neurons mimics most of the clinical (e.g. progressive paralysis) and pathological (e.g. astrocytosis and signs of axonopathy) traits of ALS [54]. Several groups have proved that TDP43 and FUS/TLS are involved in multilayered steps of RNA processing and miRNA biogenesis pathway. It has been investigated recently they have role in miRNA-related ALS [59–62]. FUS, nuclear factor complexes with Drosha along with DGCR8. This association is indispensible in miRNA processing in nucleus. Cytoplasmic TDP-43 associates with Dicer, containing TRBP. This interaction facilitates the processing of the specific pre-miRNAs by Dicer, generally a subset of the pre-miRNAs whose yield in the nucleus is regulated by TDP-43, by direct binding to their terminal loops. It was proved that TDP-43 promotes neuronal outgrowth by facilitating miRNA production. Further studies show that TDP-43 has its role in both nucleus and cytoplasm. It associates with DGCR8 containing nuclear Drosha complex in RNA-dependent and RNA-independent fashion. In nucleus, TDP-43 binds to a selected pri-miRNAs, and in cytoplasm, it binds to terminal loops of pre-miRNAs. TDP-43 facilitates neuronal outgrowth through the regulation of miRNA processing [60].
Tissue/cells | miRNAs | Changes | Biological function |
Model |
---|---|---|---|---|
Skeletal muscle |
miR-1/206 family | ↓ | Myogenic differentiation |
TDP-43 mice |
CNS | miR-29a | ↑ | ER stress- induced cell death |
SOD1-G93A mice |
Skeletal muscle | miR-206, miR-133b | ↑ | Maintenance and repair of NMJ |
SOD1-G93A mice |
Human blood serum | miR-206, miR-106b | ↑ | ALS patients | |
Lumbar spinal cord | miR-b1336, miR-b2403, miR-sb659 | ↓ | NFL stability | sALS patients |
Lumbar spinal cord | miR-b1123, miRb2948, miR- b3265, miR-b5539, miR-sb1217*, miR- sb3998 |
↑ | NFL stability | |
epSPCs | miR-124a | ↑ | epSPC fate | SOD1-G93A mice |
epSPC fate | miR-9, miR-19a, miR-19b |
↓ | epSPC fate | |
Brain microglia |
miR-155, miR-146b , miR-22, miR-365, miR-125b, miR-214 |
↑ | Microglia activation |
SOD1-G93A mice |
CSF | miR-132-5p, miR-132-3p, miR-143-3p | ↓ | sALS patients | |
CSF | miR-143-5p, miR- 574-5p |
↑ | ||
Neurons | miR-9 | ↓ | ||
Spinal cord | miR-9 | ↑ | Neuronal differentiation |
SOD1-G93A mice |
Spinal cord | miR-124a | ↓ | Glutamate transport |
SOD1-G93A mice |
Spinal cord | miR-558, miR-16-2*, miR-146a*, miR-508-5p, miR-373*, miR-551a, miR-506, miR-518a-5p, miR-518e*, miR-890 |
↑ | NFL stability | sALS human patients |
Spinal cord | miR-624, miR-520, miR-524-5p, miR- 548a-5p, miR-606, miR-612, miR-647 miR-155, miR-17, miR-19b, miR-20a, miR-24-2*, miR- 106a, miR-142-3p, miR-142-5p, miR- 146a, miR-146b, miR-223 |
↓ | NFL stability | |
Spinal cord | miR-155, miR-17, miR-19b, miR-20a, miR-24-2*, miR-106a, miR-142-3p, miR-142-5p, miR-146a, miR-146b |
SOD1-G93A mice | ||
Spinal cord | miR-24-2*, miR- 142-3p, miR-142- 5p, miR-146a, miR- 146b, miR-155 |
Human patients | ||
Blood leukocytes | miR-338-3p | ↑ | sALS human patients | |
miR-451, miR-1275, miR-328-5P, miR- 638, miR-149, miR- 665 |
↓ | |||
Skeletal muscle |
miR-23a, miR-29b, miR-206 miR-455 |
↑ | Mitochondrial function |
Human patients |
Spinal cord microglia |
miR-544, miR-23, miR-203, miR-340 miR-146. miR-130/miR-301, miR-155, miR-27, miR-16/miR-497/miR-195*, miR-20a/miR-106b/miR-17-5p |
↑ | SOD1-G93A mice | |
Spleen monocytes |
let-7, miR-15b, miR-16, miR-27a, miR-34a, miR-132, miR-146a, miR-155, miR-223, miR-451 |
↑ | SOD1-G93A mice | |
CD14+CD16− monocytes |
miR-27a, miR-155, miR-146a, miR-32- 3p |
↑ | Human ALS patients | |
Skeletal muscle |
miR-206 | ↑ | Maintenance and repair of NMJ |
SOD1-G93A mice |
One group has reported the changes that occur in the miRNA population followed by TDP-43 knockdown in cultured cells. It was observed that
15. CNS-related miRNA dysregulations in ALS pathogenesis
miR-9, an evolutionarily conserved and multifunctional neuronal miRNA, is involved in the selection of neuronal precursors from the neuroepithelium in flies and in the specification of midbrain-hindbrain boundaries in vertebrates. Both miR-124 and miR-9 participates together in neuronal differentiation. Perturbation in their function results in the susceptibility to neuronal diseases. miR-9/miR-9* expression is significantly reduced in patients with Huntington’s disease. miR-9 and miR-132 are downregulated in Alzheimer-affected brain. miR-9 has role in ALS also. There are several miRNAs, which are specific for neural cell fate and cell cycle regulation. After inducing neuronal differentiation, miR-9, miR-124a, miR-19a and miR-19b are differentially expressed in SOD1-G93A epSPCs, while miR-9 gets upregulated in SOD1-G93A differentiated cells, with respect to WT-SOD1 and control mice. Expression analysis of the predicted miRNA targets leads to identification of a functional network of Hes1. Pten, Socs1 and Stat3 genes are important for controlling epSPC fate and showing neurogenic potential in vitro. A time-course analysis and cellular distribution pattern of miR-9 were tested. It was inferred that miR-9 expression is spatially and temporally controlled in SOD1-G93A spinal cord and there is upregulation at presymptomatic stage through early symptomatic stages, mainly in the ventral horns of grey matter, where neurodegeneration is known to occur. There is a role of miR-9 in pathogenesis in induced pluripotent stem cell differentiated into postmitotic neurons and derived from ALS patients with the TDP43 A90V and M337V mutations. In all these cases, the levels of miR-9 are decreased with respect to control neurons. In
16. Role of miR-206 in disease impediment
It was reported that miR-206, a skeletal muscle-specific miRNA, strongly induced in ALS mouse model is in coincidence with the onset of neurological symptoms. It is a modifier of disease pathogenesis. In G93A-SOD1 mice, loss of miR-206 expression does not affect disease onset apparently, but accelerates disease progression, by skeletal muscle atrophy, kyphosis and paralysis, and reduces survival. In miR-206−/−/G93A-SOD1 mice, neuromuscular junctions (NMJs) are disorganized and reinnervation of denervated muscles by motor axons is postponed in the absence of miR-206. Transcript derived from the
17. Side effects of our immune system
The immune system has a huge role in maintenance of physiological equilibrium within the CNS and in controlling neuronal cell death after adverse effects start. Butovsky et al. [65] performed a comparative analysis of miRNA expression profile of inflammatory monocytes and microglia from SOD1-G93A ALS mice to generate novel biomarkers and possible therapeutics. Spleen monocytes, Ly6Chi, are recruited to the spinal cord where they proliferate during disease progression. They have pronounced proinflammatory profile in both miRNA and gene expression profile. This type of miRNA profile was found maintained in ALS patients and is apparently unique for ALS, when compared with other neuroinflammatory diseases like multiple sclerosis [65]. De Felice et al. [75, 76] did a miRNA expression profile of circulating leukocytes in a small sample of sALS patients and detected variation of eight distinct miRNAs. miR-338-3p expression is upregulated in brain tissue from ALS patients [31]. miRNAs can circulate in cell-free forms in body fluids like serum and plasma and act as signalling molecule between cells [77, 78]. Ever-increasing evidences support the idea that serum circulating secretory vesicles, including exosomes and shedding microvesicles, can function as intercellular shuttles of RNA and miRNA [60]. Muscle-enriched miR-206 is upregulated not only in SOD1-G93A mice muscles but also there is symptomatic increase in circulation in those mice. The whole transcriptome analysis of both mRNA and miRNA of sALS fibroblasts is done, in order to confirm whether dysregulated processes in the CNS might be reproduced in cells from peripheral tissues. The microprocessor complex gene DGCR8, the gene encoding Dicer enzyme and the RISC proteins AGO1/2 are downregulated in sALS fibroblasts with consequent decrease of multiple miRNAs. This creates a new horizon for generating new diagnostic pool for ALS [21].
18. Hotspot identification
MicroRNAs can bind to the same complementary target sites or it can bind to proximally located sites adjacent to other miRNA target sites. A hotspot is a stretch of nucleotide sequence, which is prone to target of several groups of miRNAs. miRNAs are regulated and expressed spatially and temporally inside cell (Figure 5). This type of regulation and expression gives miRNAs power to play a plethora of roles in various different biological processes. All the miRNAs occupying position in “hotspot” may not regulate a gene in the same pattern. Generally, predominant miRNA competitively outcasts other miRNAs and plays as a potential repressor. There is no clear evidence of the cause behind competitive outcasting and variable effectiveness. Various researchers are trying to use different standards and parameters like the same chromosomal location for more than two miRNAs, the same orientation, phylogenetic relationships and absence of interfering transcription unit.
miRNA targets are generally predicted using miRanda and then further analysed for miRNA-prone regions in all the selected genes. Generally, a region is defined as hotspot, if it has a minimum of 10 nucleotides overlapping from the starting position and occurrence of three miRNA targets. It has been shown that there are 11 miRNA-prone regions in 35 genes associated to ALS. CYCS and MAP2K3 have one hotspot each in 3′ UTR, while single target site hotspot was in 5′ UTR in four genes (Figure 5), that is, BAX, DERL1, SLC1A2 and RAB5A. Studies have shown that MAP2K3 has two hotspots by miRNA cluster prediction. Target sites for three miRNAs, i.e. hsa-miR-107, hsa-miR-423-5p and hsa-miR-103, are congregated in another hotspot found in 3′ UTR. MAP2K3, having two such hotspots, is presumed to show more sensitivity towards gene regulation by microRNAs. The small number of hotspots makes the miRNAs more specific and stringent regulator. The variability in existence of such target site hotspots adds complexity in gene regulation [9] (Figure 5).
19. Multifarious miRNA regulation
MicroRNA-mediated gene regulation adds multilayered complexity to gene expression. Distribution of target sites over several genes of one miRNA makes the picture more intricate. In principle, one miRNA can bind to more than one gene (multiplicity), and one gene can be controlled by more than one miRNA (cooperativity) [79]. Relaxed base pairing gives rise to multiplicity property. Target sites are rarely distributed on a gene. Generally they are scattered and the range of occurrence varies in numbers in a gene [80]. Often, presence of more target sites per gene indicates efficient regulation. Studies have shown that miRNAs, related to ALS, have multiple targets. After calculating multiplicity and cooperativity, scientists have shown that hsa-miR-370 showed maximum multiplicity as it showed 65 interactions with 30 gene sequences while no target site could be predicted in five genes, namely,
20. Complex interrelationship
Figure 6 conveys interaction map of miRNA and selected 35 genes. Interactions amongst genes and miRNAs are depicted with arrows, where 1 = hsa-miR-370, 2 = hsa-miR-874, 3 = hsa-miR-423-3p, 4 = hsa-miR-323-5p, 5 = hsa-miR-760, 6 = hsa-miR-149, 7 = hsa-miR-139-3p, 8 = hsa-miR-744, 9 = hsa-miR-324-3p, 10 = hsa-miR-339-3p and 11 = hsa-miR-654-5p. From Figure 6 we can understand the multilayered regulation [9].
21. Usage of miRNAs in therapeutics
The human mind is not satisfied to know the causative factors of a disease. Our main target is to reduce ailment and to do that we need good diagnostic system along with good therapeutics. miRNAs are one of a kind. Being a regulator of gene expression, it has far-reaching effect on our physiology. They are dysregulated in several diseases. Researchers are now trying to generate therapeutic targets to treat ALS. Presently, riluzole is the only FDA-approved drug to treat ALS [81] and it modestly slows the disease progression. A phase I clinical trial of an antisense oligonucleotide [ISIS 333611] has been proven effective. This oligonucleotide targets SOD1 mRNA and represses the production of mutant SOD1 and this is effective when delivered to the CSF of patients with fALS [82]. The same kind of drug is there which targets the sense strand of the C9orf72 hexanucleotide repeat and reduces the toxicity by suppressing RNA foci formation both in vivo and in vitro [30, 33, 83]. Several delivery systems are being invented day in and day out to deliver miRNA formulations and drugs to the CNS through blood-brain barrier. The first miRNA-based therapeutic is the miR-122 antagonist SPC3649, which is currently being evaluated in phase II clinical trials. It targets the hepatitis C virus. So far, this agent has not exhibited any adverse effects.
There are two basic approaches of miRNA-based therapeutics. One is miRNA antagonists which impede endogenous miRNAs that have a harmful gain of function in diseased tissues and involve the use of an anti-miR – a chemically modified antisense RNA – to knockdown miRNA. In one of the first endeavours to use antagomirs in ALS, delivery of anti-miR-155 to
It is really difficult to dissect all the pathways, which are regulated by one miRNA. Before releasing drugs in clinics, there should be several clinical trials to eliminate any deleterious effect. Williams and group [85] showed that miRNA-206 delayed ALS progression and promoted regeneration of neuromuscular synapses effectively in mice. This group investigated pathological modifications in motor axons and nerve terminals that precede motor neuron degeneration and clinical symptoms and the role of the skeletal muscle-specific miR-206 in motor neuron-skeletal muscle fibre signalling. This miRNA is significantly upregulated in SOD1-G93A mouse model of ALS. The genetic ablation of miR-206 in these ALS mice accelerates the disease progression. miR26 might be needed for compensatory regeneration of neuromuscular synapses after acute nerve injury and thus it decelerates ALS progression. miR-206 represses the translation of HDAC4 mRNA and counteracts its negative influence. This miRNA is also involved in neuromuscular gene expression and synapse formation. miR-206 decelerates ALS progression by detecting motor neuron injury and boosting compensatory regeneration of neuromuscular synapses by specifically inhibiting HDAC4 protein synthesis. HDAC4 has a deleterious role in ALS patients and it has been confirmed. It was shown in experiments that low progression rate in patients with ALS is associated to greater compensatory reinnervation and low HDAC4 levels. miR-133b is present in the same transcript encoding miR206, which is upregulated after denervation in the NMJs. The genetic ablation of miR-133b was studied in SOD1-G93A mice and it was noticed that the lack of miR-133b did not modify NMJ development or reinnervation after nerve injury and, overall, disease progression was not affected. miR-206 plays an important role in NMJ reinnervation specifically. miR-155 expression can be efficiently inhibited by anti-miR delivery to the CNS and periphery of SOD1-G93A mouse [86]. Inhibition of miR-155 can improve disease progression in ALS. Single intracerebro-ventricular injection of miR-29a-specific antagomir inhibiting CNS-related miR-29a expression in CNS can prolong the survival of ALS patients, thus suggesting miR-29a as a possible marker for disease progression. Anti-miR can be used as therapeutics in treating ALS but we should have an eye on the bystander effects [21].
22. Conclusion
Molecular neuropathology, genomics and proteomics related to it have made a real progress in the last few decades. Candidate genes and their involvement in molecular interactions and networks have opened new venues in therapeutics. ALS is polygenic and non-cell-autonomous disease. Due to the presence of many gene variants in this disease, it is difficult to predict the susceptible individuals. There is no such inheritance pattern. As miRNA population is dysregulated in ALS, candidate miRNAs can be used as biomarker and targets of therapeutics. Discoveries of ALS-related miRNAs are leading us to solve the intricate cellular network-related details. As of for now, we have drugs to slow down ALS. But we still have nothing to treat this disease completely. Intricacies in miRNA network are making the development of drug more difficult because of bystander effects. There are several scientist groups around the world trying to find out the possible therapeutics.
Truly, present write-ups address many milestones of biological consequences of microRNA that shed potential light on the in-depth mechanisms of ALS. It definitely explains the motor neuron development, thereby expanding our new knowledge of ALS post-transcriptionally. Apart from that, present knowledge laid a major foundation for development of therapeutic agents of ALS. Furthermore, covering specific miRNA signatures targeted by different genes of ALS are important for broad classification of ALS symptoms. This sectorization of microRNA expression might be important to improve the subclassification of ALS genes which provide a unique instructive role for therapeutic approaches.
References
- 1.
Howland DS, Liu J, She Y, Goad B, Maragakis NJ, Kim B, et al. Focal loss of the glutamate transporter EAAT2 in a transgenic rat model of SOD1 mutant-mediated amyotrophic lateral sclerosis [ALS]. Proc Natl Acad Sci U S A. 2002;99[3]:1604–9. - 2.
Bruijn LI, Miller TM, Cleveland DW. Unraveling the mechanisms involved in motor neuron degeneration in ALS. Annu Rev Neurosci. 2004;27:723–49. - 3.
Shabalina SA, Koonin EV. Origins and evolution of eukaryotic RNA interference. Trends Ecol Evol. 2008;23[10]:578–87. - 4.
Martens-Uzunova ES, Olvedy M, Jenster G. Beyond microRNA – novel RNAs derived from small non-coding RNA and their implication in cancer. Cancer Lett. 2013;340[2]:201–11. - 5.
Davis BN, Hata A. Regulation of microRNA biogenesis: a miRiad of mechanisms. Cell Commun Signal. 2009;7:18. - 6.
Riedmann LT, Schwentner R. miRNA, siRNA, piRNA and argonautes: news in small matters. RNA Biol. 2010;7[2]:133–9. - 7.
Jacobsen A, Silber J, Harinath G, Huse JT, Schultz N, Sander C. Analysis of microRNA-target interactions across diverse cancer types. Nat Struct Mol Biol. 2013;20[11]:1325–32. - 8.
Eulalio A, Huntzinger E, Izaurralde E. Getting to the root of miRNA-mediated gene silencing. Cell. 2008;132[1]:9–14. - 9.
Shinde S, Arora N, Bhadra U. A complex network of microRNAs expressed in brain and genes associated with amyotrophic lateral sclerosis. Int J Genomics. 2013;2013:383024. - 10.
Smrt RD, Zhao X. Epigenetic regulation of neuronal dendrite and dendritic spine development. Front Biol [Beijing]. 2010;5[4]:304–23. - 11.
Alvarez JP, Pekker I, Goldshmidt A, Blum E, Amsellem Z, Eshed Y. Endogenous and synthetic microRNAs stimulate simultaneous, efficient, and localized regulation of multiple targets in diverse species. Plant Cell. 2006;18[5]:1134–51. - 12.
Harries LW. MicroRNAs as mediators of the ageing process. Genes [Basel]. 2014;5[3]:656–70. - 13.
Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 1993;75[5]:843–54. - 14.
Reinhart BJ, Slack FJ, Basson M, Pasquinelli AE, Bettinger JC, Rougvie AE, et al. The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans . Nature. 2000;403[6772]:901–6. - 15.
Weaver R. Molecular Biology. McGraw-Hill Education; New York, NY10020, USA, 2011. - 16.
Xing L, Kieff E. cis-Acting effects on RNA processing and Drosha cleavage prevent Epstein-Barr virus latency III BHRF1 expression. J Virol. 2011;85[17]:8929–39. - 17.
Lee Y, Kim M, Han J, Yeom KH, Lee S, Baek SH, et al. MicroRNA genes are transcribed by RNA polymerase II. EMBO J. 2004;23[20]:4051–60. - 18.
Valencia-Sanchez MA, Liu J, Hannon GJ, Parker R. Control of translation and mRNA degradation by miRNAs and siRNAs. Genes Dev. 2006;20[5]:515–24. - 19.
Bartel DP. MicroRNAs: target recognition and regulatory functions. Cell. 2009;136[2]:215–33. - 20.
Hibio N, Hino K, Shimizu E, Nagata Y, Ui-Tei K. Stability of miRNA 5′ terminal and seed regions is correlated with experimentally observed miRNA-mediated silencing efficacy. Sci Rep. 2012;2:996. - 21.
Volonte C, Apolloni S, Parisi C. MicroRNAs: newcomers into the ALS picture. CNS Neurol Disord Drug Targets. 2015;14[2]:194–207. - 22.
Pillai RS. MicroRNA function: multiple mechanisms for a tiny RNA? RNA. 2005;11[12]:1753–61. - 23.
Bentwich I, Avniel A, Karov Y, Aharonov R, Gilad S, Barad O, et al. Identification of hundreds of conserved and nonconserved human microRNAs. Nat Genet. 2005;37[7]:766–70. - 24.
Winter J, Jung S, Keller S, Gregory RI, Diederichs S. Many roads to maturity: microRNA biogenesis pathways and their regulation. Nat Cell Biol. 2009;11[3]:228–34. - 25.
Tomari Y, Zamore PD. MicroRNA biogenesis: drosha can't cut it without a partner. Curr Biol. 2005;15[2]:R61–4. - 26.
Kiriakidou M, Tan GS, Lamprinaki S, De Planell-Saguer M, Nelson PT, Mourelatos Z. An mRNA m7G cap binding-like motif within human Ago2 represses translation. Cell. 2007;129[6]:1141–51. - 27.
Chendrimada TP, Finn KJ, Ji X, Baillat D, Gregory RI, Liebhaber SA, et al. MicroRNA silencing through RISC recruitment of eIF6. Nature. 2007;447[7146]:823–8. - 28.
Chan SP, Slack FJ. microRNA-mediated silencing inside P-bodies. RNA Biol. 2006;3[3]:97–100. - 29.
Duursma AM, Kedde M, Schrier M, le Sage C, Agami R. miR-148 targets human DNMT3b protein coding region. RNA. 2008;14[5]:872–7. - 30.
Renton AE, Majounie E, Waite A, Simon-Sanchez J, Rollinson S, Gibbs JR, et al. A hexanucleotide repeat expansion in C9ORF72 is the cause of chromosome 9p21-linked ALS-FTD. Neuron. 2011;72[2]:257–68. - 31.
Cortez MA, Bueso-Ramos C, Ferdin J, Lopez-Berestein G, Sood AK, Calin GA. MicroRNAs in body fluids – the mix of hormones and biomarkers. Nat Rev Clin Oncol. 2011;8[8]:467–77. - 32.
Andersen PM, Al-Chalabi A. Clinical genetics of amyotrophic lateral sclerosis: what do we really know? Nat Rev Neurol. 2011;7[11]:603–15. - 33.
DeJesus-Hernandez M, Mackenzie IR, Boeve BF, Boxer AL, Baker M, Rutherford NJ, et al. Expanded GGGGCC hexanucleotide repeat in noncoding region of C9ORF72 causes chromosome 9p-linked FTD and ALS. Neuron. 2011;72[2]:245–56. - 34.
Sreedharan J, Brown RH, Jr. Amyotrophic lateral sclerosis: problems and prospects. Ann Neurol. 2013;74[3]:309–16. - 35.
Yacila G, Sari Y. Potential therapeutic drugs and methods for the treatment of amyotrophic lateral sclerosis. Curr Med Chem. 2014;21[31]:3583–93. - 36.
Appel SH, Beers DR, Henkel JS. T cell-microglial dialogue in Parkinson's disease and amyotrophic lateral sclerosis: are we listening? Trends Immunol. 2010;31[1]:7–17. - 37.
Volonte C, Apolloni S, Carri MT, D'Ambrosi N. ALS: focus on purinergic signalling. Pharmacol Ther. 2011;132[1]:111–22. - 38.
Hossaini M, Cardona Cano S, van Dis V, Haasdijk ED, Hoogenraad CC, Holstege JC, et al. Spinal inhibitory interneuron pathology follows motor neuron degeneration independent of glial mutant superoxide dismutase 1 expression in SOD1-ALS mice. J Neuropathol Exp Neurol. 2011;70[8]:662–77. - 39.
McGown A, McDearmid JR, Panagiotaki N, Tong H, Al Mashhadi S, Redhead N, et al. Early interneuron dysfunction in ALS: insights from a mutant sod1 zebrafish model. Ann Neurol. 2013;73[2]:246–58. - 40.
Chen K, Northington FJ, Martin LJ. Inducible nitric oxide synthase is present in motor neuron mitochondria and Schwann cells and contributes to disease mechanisms in ALS mice. Brain Struct Funct. 2010;214[2–3]:219–34. - 41.
Clement AM, Nguyen MD, Roberts EA, Garcia ML, Boillee S, Rule M, et al. Wild-type nonneuronal cells extend survival of SOD1 mutant motor neurons in ALS mice. Science. 2003;302[5642]:113–7. - 42.
Ilieva H, Polymenidou M, Cleveland DW. Non-cell autonomous toxicity in neurodegenerative disorders: ALS and beyond. J Cell Biol. 2009;187[6]:761–72. - 43.
Boillee S, Vande Velde C, Cleveland DW. ALS: a disease of motor neurons and their nonneuronal neighbors. Neuron. 2006;52[1]:39–59. - 44.
Di Giorgio FP, Carrasco MA, Siao MC, Maniatis T, Eggan K. Non-cell autonomous effect of glia on motor neurons in an embryonic stem cell-based ALS model. Nat Neurosci. 2007;10[5]:608–14. - 45.
Castillo K, Nassif M, Valenzuela V, Rojas F, Matus S, Mercado G, et al. Trehalose delays the progression of amyotrophic lateral sclerosis by enhancing autophagy in motoneurons. Autophagy. 2013;9[9]:1308–20. - 46.
Vargas MR, Pehar M, Cassina P, Beckman JS, Barbeito L. Increased glutathione biosynthesis by Nrf2 activation in astrocytes prevents p75NTR-dependent motor neuron apoptosis. J Neurochem. 2006;97[3]:687–96. - 47.
Haidet-Phillips AM, Hester ME, Miranda CJ, Meyer K, Braun L, Frakes A, et al. Astrocytes from familial and sporadic ALS patients are toxic to motor neurons. Nat Biotechnol. 2011;29[9]:824–8. - 48.
Eacker SM, Dawson TM, Dawson VL. Understanding microRNAs in neurodegeneration. Nat Rev Neurosci. 2009 Dec;10[12]:837–41. - 49.
Lau P, de Strooper B. Dysregulated microRNAs in neurodegenerative disorders. Semin Cell Dev Biol. 2010;21[7]:768–73. - 50.
Sonntag KC. MicroRNAs and deregulated gene expression networks in neurodegeneration. Brain Res. 2010;1338:48–57. - 51.
Schaefer A, O'Carroll D, Tan CL, Hillman D, Sugimori M, Llinas R, et al. Cerebellar neurodegeneration in the absence of microRNAs. J Exp Med. 2007;204[7]:1553–8. - 52.
Cuellar TL, Davis TH, Nelson PT, Loeb GB, Harfe BD, Ullian E, et al. Dicer loss in striatal neurons produces behavioral and neuroanatomical phenotypes in the absence of neurodegeneration. Proc Natl Acad Sci U S A. 2008;105[14]:5614–9. - 53.
Xu Z, Yang C. TDP-43-The key to understanding amyotrophic lateral sclerosis. Rare Dis. 2014;2[1]:e944443. - 54.
Haramati S, Chapnik E, Sztainberg Y, Eilam R, Zwang R, Gershoni N, et al. miRNA malfunction causes spinal motor neuron disease. Proc Natl Acad Sci U S A. 2010;107[29]:13111–6. - 55.
Shin D, Shin JY, McManus MT, Ptacek LJ, Fu YH. Dicer ablation in oligodendrocytes provokes neuronal impairment in mice. Ann Neurol. 2009;66[6]:843–57. - 56.
Tao J, Wu H, Lin Q, Wei W, Lu XH, Cantle JP, et al. Deletion of astroglial Dicer causes non-cell-autonomous neuronal dysfunction and degeneration. J Neurosci. 2011;31[22]:8306–19. - 57.
Pereira JA, Baumann R, Norrmen C, Somandin C, Miehe M, Jacob C, et al. Dicer in Schwann cells is required for myelination and axonal integrity. J Neurosci. 2010;30[19]:6763–75. - 58.
Verrier JD, Semple-Rowland S, Madorsky I, Papin JE, Notterpek L. Reduction of Dicer impairs Schwann cell differentiation and myelination. J Neurosci Res. 2010;88[12]:2558–68. - 59.
Gregory RI, Yan KP, Amuthan G, Chendrimada T, Doratotaj B, Cooch N, et al. The microprocessor complex mediates the genesis of microRNAs. Nature. 2004;432[7014]:235–40. - 60.
Kawahara Y, Mieda-Sato A. TDP-43 promotes microRNA biogenesis as a component of the Drosha and Dicer complexes. Proc Natl Acad Sci U S A. 2012;109[9]:3347–52. - 61.
Buratti E, De Conti L, Stuani C, Romano M, Baralle M, Baralle F. Nuclear factor TDP-43 can affect selected microRNA levels. FEBS J. 2010;277[10]:2268–81. - 62.
Ling SC, Albuquerque CP, Han JS, Lagier-Tourenne C, Tokunaga S, Zhou H, et al. ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS. Proc Natl Acad Sci U S A. 2010;107[30]:13318–23. - 63.
Chen H, Qian K, Du Z, Cao J, Petersen A, Liu H, et al. Modeling ALS with iPSCs reveals that mutant SOD1 misregulates neurofilament balance in motor neurons. Cell Stem Cell. 2014;14[6]:796–809. - 64.
Tortelli R, Ruggieri M, Cortese R, D'Errico E, Capozzo R, Leo A, et al. Elevated cerebrospinal fluid neurofilament light levels in patients with amyotrophic lateral sclerosis: a possible marker of disease severity and progression. Eur J Neurol. 2012;19[12]:1561–7. - 65.
Butovsky O, Siddiqui S, Gabriely G, Lanser AJ, Dake B, Murugaiyan G, et al. Modulating inflammatory monocytes with a unique microRNA gene signature ameliorates murine ALS. J Clin Invest. 2012;122[9]:3063–87. - 66.
Ishtiaq M, Campos-Melo D, Volkening K, Strong MJ. Analysis of novel NEFL mRNA targeting microRNAs in amyotrophic lateral sclerosis. PLoS One. 2014;9[1]:e85653. - 67.
Parisi C, Arisi I, D'Ambrosi N, Storti AE, Brandi R, D'Onofrio M, et al. Dysregulated microRNAs in amyotrophic lateral sclerosis microglia modulate genes linked to neuroinflammation. Cell Death Dis. 2013;4:e959. - 68.
Chiu IM, Chen A, Zheng Y, Kosaras B, Tsiftsoglou SA, Vartanian TK, et al. T lymphocytes potentiate endogenous neuroprotective inflammation in a mouse model of ALS. Proc Natl Acad Sci U S A. 2008;105[46]:17913–8. - 69.
D'Ambrosi N, Finocchi P, Apolloni S, Cozzolino M, Ferri A, Padovano V, et al. The proinflammatory action of microglial P2 receptors is enhanced in SOD1 models for amyotrophic lateral sclerosis. J Immunol. 2009;183[7]:4648–56. - 70.
Poloni M, Facchetti D, Mai R, Micheli A, Agnoletti L, Francolini G, et al. Circulating levels of tumour necrosis factor-alpha and its soluble receptors are increased in the blood of patients with amyotrophic lateral sclerosis. Neurosci Lett. 2000;287[3]:211–4. - 71.
Hensley K, Fedynyshyn J, Ferrell S, Floyd RA, Gordon B, Grammas P, et al. Message and protein-level elevation of tumor necrosis factor alpha [TNF alpha] and TNF alpha-modulating cytokines in spinal cords of the G93A-SOD1 mouse model for amyotrophic lateral sclerosis. Neurobiol Dis. 2003;14[1]:74–80. - 72.
Foran E, Trotti D. Glutamate transporters and the excitotoxic path to motor neuron degeneration in amyotrophic lateral sclerosis. Antioxid Redox Signal. 2009;11[7]:1587–602. - 73.
Heath PR, Shaw PJ. Update on the glutamatergic neurotransmitter system and the role of excitotoxicity in amyotrophic lateral sclerosis. Muscle Nerve. 2002;26[4]:438–58. - 74.
Russell AP, Wada S, Vergani L, Hock MB, Lamon S, Leger B, et al. Disruption of skeletal muscle mitochondrial network genes and miRNAs in amyotrophic lateral sclerosis. Neurobiol Dis. 2013;49:107–17. - 75.
De Felice B, Guida M, Guida M, Coppola C, De Mieri G, Cotrufo R. A miRNA signature in leukocytes from sporadic amyotrophic lateral sclerosis. Gene. 2012;508[1]:35–40. - 76.
Shioya M, Obayashi S, Tabunoki H, Arima K, Saito Y, Ishida T, et al. Aberrant microRNA expression in the brains of neurodegenerative diseases: miR-29a decreased in Alzheimer disease brains targets neurone navigator 3. Neuropathol Appl Neurobiol. 2010;36[4]:320–30. - 77.
Reid G, Kirschner MB, van Zandwijk N. Circulating microRNAs: association with disease and potential use as biomarkers. Crit Rev Oncol Hematol. 2011;80[2]:193–208. - 78.
Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 2007;9[6]:654–9. - 79.
Enright AJ, John B, Gaul U, Tuschl T, Sander C, Marks DS. MicroRNA targets in Drosophila. Genome Biol. 2003;5[1]:R1. - 80.
John B, Enright AJ, Aravin A, Tuschl T, Sander C, Marks DS. Human microRNA targets. PLoS Biol. 2004;2[11]:e363. - 81.
King IN, Yartseva V, Salas D, Kumar A, Heidersbach A, Ando DM, et al. The RNA-binding protein TDP-43 selectively disrupts microRNA-1/206 incorporation into the RNA-induced silencing complex. J Biol Chem. 2014;289[20]:14263–71. - 82.
Miller TM, Pestronk A, David W, Rothstein J, Simpson E, Appel SH, et al. An antisense oligonucleotide against SOD1 delivered intrathecally for patients with SOD1 familial amyotrophic lateral sclerosis: a phase 1, randomised, first-in-man study. Lancet Neurol. 2013;12[5]:435–42. - 83.
Lee YB, Chen HJ, Peres JN, Gomez-Deza J, Attig J, Stalekar M, et al. Hexanucleotide repeats in ALS/FTD form length-dependent RNA foci, sequester RNA binding proteins, and are neurotoxic. Cell Rep. 2013;5[5]:1178–86. - 84.
Paez-Colasante X, Figueroa-Romero C, Sakowski SA, Goutman SA, Feldman EL. Amyotrophic lateral sclerosis: mechanisms and therapeutics in the epigenomic era. Nat Rev Neurol. 2015;11[5]:266–79. - 85.
Williams AH, Valdez G, Moresi V, Qi X, McAnally J, Elliott JL, et al. MicroRNA-206 delays ALS progression and promotes regeneration of neuromuscular synapses in mice. Science. 2009;326[5959]:1549–54. - 86.
Koval ED, Shaner C, Zhang P, du Maine X, Fischer K, Tay J, et al. Method for widespread microRNA-155 inhibition prolongs survival in ALS-model mice. Hum Mol Genet. 2013;22[20]:4127–35.