Blocking a MicroRNA Slows Motor Neuron Disease in Mice

Series - Society for Neuroscience Annual Meeting 2013:

Researchers are taking aim at amyotrophic lateral sclerosis with an antisense oligonucleotide targeted toward microRNA (miRNA), those nucleic acid snippets that each regulate hundreds of mRNAs. A single anti-miRNA therapeutic could tune a whole network of genes, suggests Erica Koval of Washington University in St. Louis. She presented preclinical data on anti-miR-155 at “RNA Metabolism in Neurological Disease,” a meeting held November 7–8 in San Diego. Koval reported that blocking miR-155 in mouse models of ALS slowed disease progression. Koval, senior author Timothy Miller, and colleagues published some of the findings last month (Koval et al., 2013).

Anti-microRNA therapy has been tested for conditions including hepatitis, stroke, and cancer (Janssen et al., 2013; Buller et al., 2010; Abba et al., 2012), but Koval believes she is the first to try it for a neurodegenerative condition. Anti-miRNA oligonucleotides work a bit differently than standard antisense molecules aimed at much larger mRNAs. While the latter destroy the mRNA target, anti-microRNAs simply bind to their targets, preventing them from interacting with the mRNAs they regulate. Koval and Miller collaborated with Regulus Therapeutics of San Diego, a spinoff of Isis Pharmaceuticals of Carlsbad, California. Isis develops traditional anti-mRNA treatments, including an antisense therapy for the ALS gene superoxide dismutase 1 (SOD1) in collaboration with Miller and others (see May 2013 news story). Regulus focuses on anti-microRNAs.

At the start of the project, Koval and colleagues predicted that microRNAs would be dysregulated in ALS, and that correcting the dysregulation might help. Using microarrays, she identified 12 microRNAs that were upregulated in mouse and rat models of ALS, compared to control animals. Of these, six were also upregulated in the spinal cords of people who had died of ALS, according to Koval’s analysis of autopsy tissues. No microRNAs were significantly downregulated. Koval chose to focus on miR-155 first, because levels were doubled in spinal cords from people who had either familial or sporadic ALS, indicating a common problem. Researchers already knew that MiR-155 promotes neuroinflammation (O’Connell et al., 2010), a key feature of ALS pathology (see Boillée et al., 2006; Sep 2008 news story), and Regulus already had an antisense molecule ready to evaluate in animals.

MiR-155’s targets include mRNAs for these genes:

  • suppressor of cytokine signaling-1 (SOCS-1), which inhibits cytokine production and promotes interferon-γ activity (Wang et al., 2009);
  • cysteine-rich protein 61 (CYR61), which has diverse roles in cell proliferation and differentiation, angiogenesis, apoptosis, and creating extracellular matrix (Zhang et al., 2010);
  • and caspase recruitment domain family, member 11 (CARD11), which activates NF-κB (Xu et al., 2010).

Because the blood-brain barrier prevents microRNAs from entering the central nervous system (CNS), Koval delivered a constant stream of anti-miR-155 directly into mouse brain lateral ventricles using a micropump. Tracking the fluorescently labeled oligonucleotide, she saw that it entered neurons and glia throughout the brain and spinal cord of wild-type mice (see figure below). She also confirmed that the two-week treatment derepressed miR-155 target mRNAs; levels of SHIP1, PU.1, CARD11, and CYR61 mRNA all rose.

 

image

Infused into the lateral ventricles of mice, Cy5-labeled anti-miR-155 (red), infiltrated neurons (top) , microglia (middle), and astrocytes (bottom) labeled with antibodies (green) to NeuN, Iba1, and GFAP, respectively. Image courtesy of Erica Koval, Washington University in St. Louis.

“It is fantastic how well these things get into the nervous system,” commented Peter Nelson of the University of Kentucky in Lexington, who was not involved in the work. “Anti-microRNA provides such a ready opportunity for treatment.” Nelson speculated that neurons might take up the oligonucleotides by some sort of active mechanism.

Next, Koval tried the anti-miR-155 treatment in an ALS mouse model that expresses mutant human SOD1. She was uncertain if cells in the CNS, peripheral immune cells infiltrating the CNS, or both might be involved in any possible benefits. Notably, peripheral macrophages undergo activation in ALS (see Nov 2009 news story). To maximize her chances of treating the disease, Koval combined administration to the brain lateral ventricles with weekly peripheral injections. She began treatment when the mice were 60 days old, about 40 days before symptoms of disease—weight loss and paralysis—normally emerge. Disease onset was unchanged, but the treated mice lived 10 days longer than untreated controls, surviving to an average of 139 days. This may seem a short reprieve, but for mSOD1 mice it was comparable to results obtained with other treatments, such as SOD1 antisense therapy (see Jul 2006 news story).

Koval and Miller are now investigating precisely how blocking miR-155 increases lifespan. The immune system’s role in the disease has proved to be complex, with both positive and negative effects (see Sep 2009 news story; Oct 2008 news story; Henkel et al., 2009). Koval speculated that the treatment dampens inflammatory processes that would otherwise exacerbate the disease. “It is presumably just a way of treating inflammation in the nervous system,” agreed Nelson. Therefore, the treatment would be unlikely to halt disease in people but might slow it down, Miller said. While researchers cannot directly correlate life extension in mice to that in humans, Miller hopes that an anti-miR-155 therapeutic would offer more than the extra few months people get from taking riluzole, currently the only approved treatment for ALS.

The Miller lab and Regulus are not headed for the clinic just yet. Toxicology work remains, and Koval needs to determine whether peripheral or CNS treatment, or both, explains the benefits. A peripheral treatment would be more convenient in the clinic, though Koval noted that researchers are making excellent progress with intrathecal delivery of antisense treatments directly to the CNS (see Mar 2011 news story; Jan 2013 news story).—Amber Dance

Comments

  1. The authors of this interesting report have nicely demonstrated a new method of inhibiting micro RNAs (miRNAs) in vivo in a mouse model of ALS. Essentially, they have utilized oligonucleotides as miRNA inhibitors. By targeting one such miRNA, miRNA-155, they demonstrate improvement in their ALS mouse model, about a 10-day prolonged survival. From a neuroinflammation perspective, what I find most intriguing is that miRNA-155 is avidly taken up by microglia and astrocytes. While the authors do not further investigate a putative mechanism to account for this, it would be interesting to determine if an active process is at work here (e.g., fluid-phase pinocytosis) or whether some form of glial-neuronal interaction is taking place. Nonetheless, this work raises our awareness of the therapeutic potential of blocking miRNAs for neurodegenerative disease.

    View all comments by Terrence Town

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References

News Citations

  1. Paper Alert: Antisense Oligonucleotide Therapy Safe for ALS?
  2. Survivor ALS Models—Immunity Protects Against Mutant SOD
  3. Peripheral Innate Immunity—Not So Peripheral to ALS?
  4. Treatment Trends: Tapping Stem Cells, DNA, and RNA to Save Neurons
  5. ALS: T Cells Step Up
  6. Microglia in ALS: Helpful, Harmful, or Neutral?
  7. Research Brief: Researchers Solicit SMN Understudy to Treat SMA
  8. Cut to the Chase: Therapies Go Directly to Central Nervous System

Paper Citations

  1. Koval ED, Shaner C, Zhang P, du Maine X, Fischer K, Tay J, Chau BN, Wu GF, Miller TM. Method for widespread microRNA-155 inhibition prolongs survival in ALS-model mice. Hum Mol Genet. 2013 Jun 19; PubMed.
  2. Janssen HL, Reesink HW, Lawitz EJ, Zeuzem S, Rodriguez-Torres M, Patel K, van der Meer AJ, Patick AK, Chen A, Zhou Y, Persson R, King BD, Kauppinen S, Levin AA, Hodges MR. Treatment of HCV infection by targeting microRNA. N Engl J Med. 2013 May 2;368(18):1685-94. PubMed.
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  5. O'Connell RM, Kahn D, Gibson WS, Round JL, Scholz RL, Chaudhuri AA, Kahn ME, Rao DS, Baltimore D. MicroRNA-155 promotes autoimmune inflammation by enhancing inflammatory T cell development. Immunity. 2010 Oct 29;33(4):607-19. PubMed.
  6. Boillée S, Yamanaka K, Lobsiger CS, Copeland NG, Jenkins NA, Kassiotis G, Kollias G, Cleveland DW. Onset and progression in inherited ALS determined by motor neurons and microglia. Science. 2006 Jun 2;312(5778):1389-92. PubMed.
  7. O'Connell RM, Chaudhuri AA, Rao DS, Baltimore D. Inositol phosphatase SHIP1 is a primary target of miR-155. Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):7113-8. PubMed.
  8. Leung WH, Tarasenko T, Bolland S. Differential roles for the inositol phosphatase SHIP in the regulation of macrophages and lymphocytes. Immunol Res. 2009;43(1-3):243-51. PubMed.
  9. Wang X, Zhao Q, Matta R, Meng X, Liu X, Liu CG, Nelin LD, Liu Y. Inducible nitric-oxide synthase expression is regulated by mitogen-activated protein kinase phosphatase-1. J Biol Chem. 2009 Oct 2;284(40):27123-34. PubMed.
  10. Zhang Y, Diao Z, Su L, Sun H, Li R, Cui H, Hu Y. MicroRNA-155 contributes to preeclampsia by down-regulating CYR61. Am J Obstet Gynecol. 2010 May;202(5):466.e1-7. PubMed.
  11. Vigorito E, Perks KL, Abreu-Goodger C, Bunting S, Xiang Z, Kohlhaas S, Das PP, Miska EA, Rodriguez A, Bradley A, Smith KG, Rada C, Enright AJ, Toellner KM, Maclennan IC, Turner M. microRNA-155 regulates the generation of immunoglobulin class-switched plasma cells. Immunity. 2007 Dec;27(6):847-59. PubMed.
  12. Xu G, Fewell C, Taylor C, Deng N, Hedges D, Wang X, Zhang K, Lacey M, Zhang H, Yin Q, Cameron J, Lin Z, Zhu D, Flemington EK. Transcriptome and targetome analysis in MIR155 expressing cells using RNA-seq. RNA. 2010 Aug;16(8):1610-22. PubMed.
  13. Henkel JS, Beers DR, Zhao W, Appel SH. Microglia in ALS: the good, the bad, and the resting. J Neuroimmune Pharmacol. 2009 Dec;4(4):389-98. PubMed.

External Citations

  1. antisense therapy

Further Reading

Papers

  1. Zhang Z, Almeida S, Lu Y, Nishimura AL, Peng L, Sun D, Wu B, Karydas AM, Tartaglia MC, Fong JC, Miller BL, Farese RV, Moore MJ, Shaw CE, Gao FB. Downregulation of MicroRNA-9 in iPSC-Derived Neurons of FTD/ALS Patients with TDP-43 Mutations. PLoS One. 2013;8(10):e76055. PubMed.
  2. Lungu G, Stoica G, Ambrus A. MicroRNA profiling and the role of microRNA-132 in neurodegeneration using a rat model. Neurosci Lett. 2013 Oct 11;553:153-8. PubMed.
  3. Campos-Melo D, Droppelmann CA, He Z, Volkening K, Strong MJ. Altered microRNA expression profile in Amyotrophic Lateral Sclerosis: a role in the regulation of NFL mRNA levels. Mol Brain. 2013;6:26. PubMed.
  4. Lehmann SM, Krüger C, Park B, Derkow K, Rosenberger K, Baumgart J, Trimbuch T, Eom G, Hinz M, Kaul D, Habbel P, Kälin R, Franzoni E, Rybak A, Nguyen D, Veh R, Ninnemann O, Peters O, Nitsch R, Heppner FL, Golenbock D, Schott E, Ploegh HL, Wulczyn FG, Lehnardt S. An unconventional role for miRNA: let-7 activates Toll-like receptor 7 and causes neurodegeneration. Nat Neurosci. 2012 May 20; PubMed.
  5. Kawahara Y. Implications of microRNA dysfunction in the pathogenesis of ALS. Rinsho Shinkeigaku. 2010 Nov;50(11):979-81. PubMed.
  6. 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 May;277(10):2268-81. PubMed.

Primary Papers

  1. Koval ED, Shaner C, Zhang P, du Maine X, Fischer K, Tay J, Chau BN, Wu GF, Miller TM. Method for widespread microRNA-155 inhibition prolongs survival in ALS-model mice. Hum Mol Genet. 2013 Jun 19; PubMed.

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