SBIR-STTR Award

Multiple Sclerosis (MS) is the most common neurological disease of early adulthood and is mediated by autoimmune mechanisms that lead to demyelination and neuronal damage in the central nervous system, resulting in progressive neurological dysfunction. Up to date, there is no cure for the disease and current available treatments focus on preventing future immunological attacks, primarily by suppressing the immune system, and this has adverse side effects that are often severe or fatal. Accordingly, there is a clear unmet need for the development of effective and well-tolerated therapies to arrest MS development. This has been challenging because MS has multiple etiologies (>150 genes identified as risk factors for MS so far) and the molecular mechanisms underlying these etiologies are not well understood. We uncovered the molecular underpinnings of an MS etiology and hope that this knowledge will translate into an accurate therapy for MS. The etiology in question is associated with the interleukin 7 receptor (IL7R) gene, which encodes a cell surface receptor in T cells (hereafter referred to as mIL7R) that plays a central role in the homeostasis of T cells. We previously identified the genetic variant rs6897932 within exon 6 of IL7R to be strongly associated with increased MS risk. Furthermore, we showed that the risk allele of this variant increases exclusion (skipping) of the alternative exon 6 leading to a higher fraction of mRNAs encoding a secreted form of the receptor (sIL7R), leading to elevated levels of circulating sIL7R. This has important repercussions in the development of MS because sIL7R has been shown to aggravate the progression and severity of the disease in the Experimental Autoimmune Encephalomyelitis (EAE) mouse model of MS. Here, we propose to develop a novel biologic MS drug centered on preventing formation of the pathogenic sIL7R. Given that sIL7R is produced by exclusion of exon 6 from IL7R mRNAs, our strategy is to generate splicing-modulating antisense oligonucleotides (ASOs) to correct splicing of IL7R exon 6 and restore expression of IL7R protein isoforms. Our approach represents a major improvement over current MS therapies in that by correcting IL7R splicing, it will diminish expression of the pathogenic sIL7R isoform, without reducing expression of the mIL7R. This is important because mIL7R function is essential for proper immune function and its disruption leads to immunodeficiency. Therefore, our biologic drug, unlike current MS drugs, will not cause immunosuppression. Further supporting a role of IL7R in the development of autoimmunity, this gene has been associated with other autoimmune diseases including Type I Diabetes (T1D), Rheumatoid Arthritis (RA) and Systemic Lupus Erythematosus (SLE), and most importantly, patients of these diseases have been shown to have elevated levels of circulating sIL7R. Accordingly, the approach proposed here has the potential to yield the first accurate therapy for MS and several autoimmune diseases associated with elevated levels of sIL7R.

Public Health Relevance Statement:
PROJECT NARRATIVE Multiple Sclerosis (MS) is a demyelinating autoimmune disorder of the central nervous system that causes progressive neurological dysfunction and disability in young adults, and this represents a major socio- economical burden for the patients and society. There is no curative treatment for this devastating disease and the current drugs, while providing hope, are far from ideal as they are immune-modulators that cause adverse side effects associated with broad immunosuppression. The research proposed here aims to develop an effective and safer accurate MS therapy that avoids immunosuppression, which is a critical unmet need in MS.

Project Terms:
Alternative Splicing; Alternative RNA Splicing; Alternate Splicing; Animals; Rheumatoid Arthritis; rheumatic arthritis; Atrophic Arthritis; Autoimmune Diseases; autoimmune disorder; Autoimmunity; Autoimmune Status; Biological Assay; Biologic Assays; Bioassay; Assay; Callithrix; Short-Tusked Marmoset; Marmosets; Hapale; cell culture; Cell Culture Techniques; cultured cell line; Strains Cell Lines; CellLine; Cell Line; Cell Body; Cells; Cultured Cells; Central Nervous System Disorders; CNS disorder; CNS Diseases; Central Nervous System Diseases; Chemistry; Demyelinations; type one diabetes; type I diabetes; ketosis prone diabetes; juvenile diabetes mellitus; juvenile diabetes; insulin dependent diabetes; Type I Diabetes Mellitus; Type 1 diabetes; Type 1 Diabetes Mellitus; T1DM; T1D; T1 diabetes; T1 DM; Sudden-Onset Diabetes Mellitus; Ketosis-Prone Diabetes Mellitus; Juvenile-Onset Diabetes Mellitus; IDDM; Brittle Diabetes Mellitus; Insulin-Dependent Diabetes Mellitus; Diagnosis; Disorder; Disease; drug/agent; Pharmaceutic Preparations; Medication; Drugs; Pharmaceutical Preparations; pseudohypertrophic muscular paralysis; pseudohypertrophic adult muscular dystrophy; progressive muscular dystrophy of childhood; mild X-linked recessive muscular dystrophy; classic X-linked recessive muscular dystrophy; childhood pseudohypertrophic muscular dystrophy; benign X-linked recessive muscular dystrophy; X-linked recessive muscular dystrophy; X-linked muscular dystrophy; X-linked dilated cardiomyopathy; Pseudohypertrophic Muscular Dystrophy; Ellis-van Creveld (EvC) syndrome; Duchenne-Griesinger syndrome; Duchenne; Duchene; Duchenne muscular dystrophy; autoimmune encephalomyelitis; Experimental Autoimmune Encephalitis; Experimental Allergic Encephalomyelitis; Experimental Allergic Encephalitis; EAE; Experimental Autoimmune Encephalomyelitis; Exons; Future; Genes; Goals; Physiological Homeostasis; Autoregulation; Homeostasis; allergic/immunologic organ system; allergic/immunologic body system; Immune system; immunodeficiency; immune deficiency disorder; hypoimmunity; Immunological Deficiency Syndromes; Immunodeficiency Syndrome; Immunodeficiency Disorder; Immunologic Deficiency Syndromes; immune suppression; Immunosuppressive Effect; Immunosuppression Effect; Immunosuppression; immuno therapy; immune-based treatments; immune-based therapies; immune therapy; immune therapeutic strategy; immune therapeutic regimens; immune therapeutic interventions; immune therapeutic approach; immune drugs; Immunologically Directed Therapy; Immunotherapy; In Vitro; interferon beta 2; Plasmacytoma Growth Factor; Myeloid Differentiation-Inducing Protein; MGI-2; IL6 Protein; IL-6; IFNB2; IFN-beta 2; Hybridoma Growth Factor; Hepatocyte-Stimulating Factor; HPGF; BSF2; BSF-2; BCDF; B-Cell Stimulatory Factor-2; B-Cell Differentiation Factor-2; B-Cell Differentiation Factor; B cell stimulating factor 2; B cell differentiation factor; Interleukin-6; renal; Kidney Urinary System; Kidney; heavy metal lead; heavy metal Pb; Pb element; Lead; systemic lupus erythematosis; disseminated lupus erythematosus; Systemic SLE - Lupus Erythematosus; Systemic Lupus Erythmatosus; SLE; Lupus Erythematosus Disseminatus; Systemic Lupus Erythematosus; Macaque; Macaca; insular sclerosis; Disseminated Sclerosis; Multiple Sclerosis; Murine; Mice Mammals; Mice; Mus; Cruveilhier disease; Aran-Duchenne disease; Spinal Muscular Atrophy; neurological disease; Neurological Disorders; Neurologic Disorders; Nervous System Diseases; nervous system disorder; neuronal; Neurocyte; Neural Cell; Nerve Unit; Nerve Cells; Neurons; living system; Organism; Patients; Play; Cell Surface Receptors; Research; Risk; Risk Factors; Splicing; RNA Splicing; mRNA; Messenger RNA; social role; Role; Safety; Societies; thymus derived lymphocyte; T-Cells; T-Lymphocyte; T4 Lymphocytes; T4 Cells; CD4-Positive Lymphocytes; CD4+ T-Lymphocyte; CD4 lymphocyte; CD4 helper T cell; CD4 T cells; CD4 Cells; CD4 Positive T Lymphocytes; Testing; Translating; United States; Work; antisense oligo; anti-sense oligo; anti-sense agent; Antisense Agent; Anti-Sense Oligonucleotides; Antisense Oligonucleotides; Interleukin 7 Receptor Alpha; IL7 Receptors; IL-7 Receptors; CD127 Antigens; Interleukin 7 Receptor; Mediating; medical costs; Medical Care Costs; base; improved; Phase; Biological; Medical; disability; nonhuman primate; non-human primate; young adult; young adulthood; adult youth; Therapeutic; Genetic; Reporter; Knowledge; Immune; Immunes; Complex; Clinic; Autoimmune Process; Autoimmune; Sensory; cell type; Severity of illness; disease severity; Protein Isoforms; Isoforms; receptor; Receptor Protein; receptor expression; success; neuron loss; neuronal loss; neuronal death; neuronal cell loss; neuronal cell death; neuron death; neuron cell loss; neuron cell death; nerve cell loss; nerve cell death; Animal Model; model organism; model of animal; Animal Models and Related Studies; cis acting element; Toxic effect; Toxicities; novel; Exclusion; Neurologic Dysfunctions; neurological dysfunction; Therapeutic Intervention; intervention therapy; Emotional; Pharmacodynamics; high throughput screening; High Throughput Assay; receptor function; Neuraxis; Central Nervous System; CNS Nervous System; Receptor Up-Regulation; Pathogenicity; Bioinformatics; Bio-Informatics; Cellular Immune Function; immune function; preventing; prevent; disease causation; causation; Causality; Etiology; Receptor Gene; Molecular Analysis; in vivo; in vivo Model; Cognitive; Transcript; Immunologics; Immunologically; Immunological; Immunologic; Immunochemical Immunologic; Molecular; Modification; Development; developmental; safety study; genetic variant; genomic variant; allelic variant; allele variant; Gene variant; pharmacokinetic model; mouse model; murine model; motor impairment; movement limitation; movement impairment; emerging adult; early adulthood; risk variant; risk locus; risk loci; risk genotype; risk gene; risk allele; Risk-associated variant; multiple sclerosis patient; patients with multiple sclerosis; patients with MS; MS patient; multiple sclerosis treatment; MS treatment; curative treatments; curative therapy; curative therapeutic; curative intervention; efficacy study; side effect

Multiple Sclerosis (MS) is the most common neurological disease of early adulthood and is mediated by autoimmune mechanisms that lead to demyelination and neuronal damage in the central nervous system, resulting in progressive neurological dysfunction. Up to date, there is no cure for this devastating disease and current available treatments focus on preventing future immunological attacks, primarily by suppressing the immune system, and this has adverse side effects that are often severe or fatal. Accordingly, there is a clear unmet need for the development of effective and well-tolerated therapies to arrest MS development. To reduce side effects, MS drugs should avoid immunosuppressive mechanisms and should be targeted to specific etiologies. This has been challenging because MS has multiple etiologies (>500 genes identified as risk factors for MS so far) and the molecular mechanisms underlying these etiologies are not well understood. Addressing this unmet need, we developed a personalized therapy that corrects a specific etiology of MS caused by elevated levels of the soluble form of the Interleukin 7 Receptor (sIL7R), which rises to pathogenic levels by aberrant exclusion of IL7R exon 6 during pre-mRNA splicing. Implicating sIL7R in the pathogenesis of MS and autoimmunity, it has been shown to: (i) be up-regulated by MS risk variants, (ii) exacerbate the severity of the disease in the Experimental Autoimmune Encephalomyelitis (EAE) mouse model of MS, and (iii) be elevated in patients from several autoimmune diseases, including MS, Type 1 diabetes, Rheumatoid arthritis and Systemic lupus erythematosus. Collectively, these findings support the scientific premise that a reduction in sIL7R would be therapeutic in MS and perhaps other autoimmune disorders where sIL7R is up-regulated. Given that sIL7R is generated by exclusion of exon 6 from IL7R RNAs, we developed antisense oligonucleotides (ASOs) that promote inclusion of this critical exon and reduce sIL7R expression (anti-sIL7R ASOs). By correcting this etiology of MS, anti-sIL7R ASOs are predicted to effectively prevent MS relapses while reducing side effects associated with immunosuppression. In our Phase I research, we optimized these ASOs ex vivo to efficiently reduce sIL7R secretion in human primary T cells with minimal cellular toxicity. In vivo efficacy studies of anti-sIL7R ASOs are limited to nonhuman primates (NHPs) because alternative splicing of IL7R exon 6 is observed exclusively in primates, of which macaques are the ideal model since they express sIL7R at levels equal to those observed in MS patients that suffer from this etiology. To advance pre-clinical development of anti-sIL7R ASOs as potential therapeutic drugs for MS, in this Phase II proposal we will assess the biodistribution, safety and therapeutic efficacy of lead anti-sIL7R ASOs in the highly relevant EAE model in cynomolgus macaques (Macaca fascicularis). This pivotal study will set the foundation for ensuing IND-enabling studies and clinical trials, and if successful, have the potential to provide a first-in-class precision medicine for MS.

Public Health Relevance Statement:
PROJECT NARRATIVE Multiple Sclerosis (MS) is a demyelinating autoimmune disorder of the central nervous system that causes progressive neurological dysfunction and disability in young adults with major socio-economical burdens for the patients and society. There is no curative treatment for this devastating disease and the current drugs, while providing hope, are far from ideal as they are immune-modulators that cause adverse side effects associated with immunosuppression. The research proposed here will test in nonhuman primates a targeted therapy for MS that avoids immunosuppressive mechanisms, which is a critical unmet need in MS.

Project Terms:
Affect; Alternative Splicing; Alternate Splicing; Alternative RNA Splicing; Animals; Rheumatoid Arthritis; Atrophic Arthritis; rheumatic arthritis; Autoimmune Diseases; autoimmune condition; autoimmune disorder; Autoimmunity; Autoimmune Status; Back; Dorsum; Biological Response Modifier Therapy; Biological Therapy; biological therapeutic; biological treatment; biologically based therapeutics; biotherapeutics; biotherapy; Brain; Brain Nervous System; Encephalon; Clinical Trials; Demyelinations; demyelinate; Insulin-Dependent Diabetes Mellitus; Brittle Diabetes Mellitus; IDDM; Juvenile-Onset Diabetes Mellitus; Ketosis-Prone Diabetes Mellitus; Sudden-Onset Diabetes Mellitus; T1 DM; T1 diabetes; T1D; T1DM; Type 1 Diabetes Mellitus; Type 1 diabetes; Type I Diabetes Mellitus; insulin dependent diabetes; juvenile diabetes; juvenile diabetes mellitus; ketosis prone diabetes; type I diabetes; type one diabetes; Disease; Disorder; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Experimental Autoimmune Encephalomyelitis; EAE; Experimental Allergic Encephalitis; Experimental Allergic Encephalomyelitis; Experimental Autoimmune Encephalitis; autoimmune encephalomyelitis; Exons; Foundations; Future; Genes; Goals; Heart; Human; Modern Man; Immune system; allergic/immunologic body system; allergic/immunologic organ system; Immunosuppression; Immunosuppression Effect; Immunosuppressive Effect; immune suppression; immune suppressive activity; immune suppressive function; immunosuppressive activity; immunosuppressive function; In Vitro; Kidney; Kidney Urinary System; renal; Lead; Pb element; heavy metal Pb; heavy metal lead; Liver; hepatic body system; hepatic organ system; Lung; Lung Respiratory System; pulmonary; Systemic Lupus Erythematosus; Lupus Erythematosus Disseminatus; SLE; Systemic Lupus Erythematous; Systemic Lupus Erythmatosus; disseminated lupus erythematosus; systemic lupus erythematosis; lymph nodes; Lymph Node Reticuloendothelial System; Lymph node proper; Lymphatic nodes; lymph gland; Relapsing-Remitting Multiple Sclerosis; Maximal Tolerated Dose; Maximally Tolerated Dose; Maximum Tolerated Dose; Modeling; cancer therapy; Cancer Treatment; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; anti-cancer therapy; anticancer therapy; cancer-directed therapy; receptor function; Neuraxis; CNS Nervous System; Central Nervous System; Pathogenicity; Cellular Immune Function; immune function; preventing; prevent; Causality; causation; disease causation; Etiology; CD127; CDW127; IL-7R alpha chain; IL-7R-alpha; IL-7Ralpha; IL-7Ra; IL7R; alpha chain interleukin-7 receptor; IL7R gene; Address; Dose; Economic Burden; Harvest; Immunodeficiency and Cancer; in vivo; Antisense Oligonucleotide Therapy; ASO therapeutics; ASO therapy; ASO treatment; anti-sense oligonucleotide therapy; anti-sense therapy; antisense therapy; Immunologics; Immunochemical Immunologic; Immunologic; Immunological; Immunologically; Monitor; Molecular; socioeconomics; socio-economic; socio-economically; socioeconomically; Development; developmental; Immunomodulators; IMiD; Immune modulatory therapeutic; immune modulating agents; immune modulating drug; immune modulating therapeutics; immune modulators; immune modulatory agents; immune modulatory drugs; immunomodulating agents; immunomodulatory agents; immunomodulatory drugs; immunomodulatory therapeutics; Image; imaging; preclinical study; pre-clinical study; Advanced Development; Biodistribution; Treatment Efficacy; intervention efficacy; therapeutic efficacy; therapy efficacy; Outcome; Coupled; Cell model; Cellular model; novel therapeutics; new drug treatments; new drugs; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel therapy; mouse model; murine model; motor impairment; movement impairment; movement limitation; emerging adult; early adulthood; efficacy testing; drug candidate; risk variant; Risk-associated variant; risk allele; risk gene; risk genotype; risk loci; risk locus; precision medicine; precision-based medicine; personalized medicine; personalization of treatment; personalized therapy; personalized treatment; targeted treatment; targeted drug therapy; targeted drug treatments; targeted therapeutic; targeted therapeutic agents; targeted therapy; multiple sclerosis patient; MS patient; patients with MS; patients with multiple sclerosis; multiple sclerosis treatment; MS treatment; multiple sclerosis therapy; curative treatments; curative intervention; curative therapeutic; curative therapy; reduce symptoms; alleviate symptom; ameliorating symptom; decrease symptom; fewer symptoms; relieves symptoms; symptom alleviation; symptom reduction; symptom relief; efficacy study; preclinical development; pre-clinical development; primary endpoint; primary end point; side effect; Immune mediated destruction; lymphnodes; Macaca; Macaque; Macaca fascicularis; Cynomolgus Monkey; Cynomolgus macaque; M fascicularis; M. fascicularis; Crab-Eating Macaque; Crab-Eating Monkey; Magnetic Resonance Imaging; MR Imaging; MR Tomography; MRI; MRIs; Medical Imaging, Magnetic Resonance / Nuclear Magnetic Resonance; NMR Imaging; NMR Tomography; Nuclear Magnetic Resonance Imaging; Zeugmatography; Morbidity - disease rate; Morbidity; mortality; mRNA Precursor; Pre-mRNA; RNA, Messenger, Precursors; premRNA; Multiple Sclerosis; Disseminated Sclerosis; insular sclerosis; Mus; Mice; Mice Mammals; Murine; Myelin Sheath; Neuron Degeneration; neural degeneration; neurodegeneration; neurodegenerative; neurological degeneration; neuronal degeneration; Nerve Degeneration; Nervous System Diseases; Neurologic Disorders; Neurological Disorders; neurological disease; nervous system disorder; Nerve Cells; Nerve Unit; Neural Cell; Neurocyte; neuronal; Neurons; Pathology; Patients; Pharmacokinetics; Drug Kinetics; Primates Mammals; Primates; Relapse; Research; Risk Factors; Non-Polyadenylated RNA; RNA Gene Products; Ribonucleic Acid; RNA; Splicing; RNA Splicing; Safety; Societies; Medulla Spinalis; Spinal Cord; Spleen Reticuloendothelial System; Spleen; T-Cells; thymus derived lymphocyte; T-Lymphocyte; CD4 Cells; CD4 T cells; CD4 helper T cell; CD4 lymphocyte; CD4+ T-Lymphocyte; CD4-Positive Lymphocytes; T4 Cells; T4 Lymphocytes; CD4 Positive T Lymphocytes; Testing; Thymus; Thymus Proper; Thymus Reticuloendothelial System; Thymus Gland; Time; Tissues; Body Tissues; Toxicology; Antisense Oligonucleotides; Anti-Sense Oligonucleotides; Antisense Agent; anti-sense agent; anti-sense oligo; antisense oligo; Interleukin 7 Receptor; CD127 Antigens; IL-7 Receptors; IL7 Receptors; Interleukin 7 Receptor Alpha; Diagnostic tests; Mediating; base; Clinical; Phase; Evaluation; disability; Individual; non-human primate; nonhuman primate; adult youth; young adulthood; young adult; Disease Progression; anti-cancer immunotherapy; anticancer immunotherapy; immune-based cancer therapies; immunotherapy for cancer; immunotherapy of cancer; cancer immunotherapy; Therapeutic; Genetic Predisposition; Genetic Susceptibility; Inherited Predisposition; Inherited Susceptibility; genetic etiology; genetic mechanism of disease; genetic vulnerability; genetically predisposed; Genetic Predisposition to Disease; Event; Autoimmune Mechanism; Autoimmune Process; disease severity; Severity of illness; Neurologic Deficit; receptor expression; cellular targeting; Toxicities; Toxic effect; Immunomodulation; immune modulation; immune regulation; immunologic reactivity control; immunomodulatory; immunoregulatory; immunoregulation; novel; Exclusion; Pathogenesis; neurological dysfunction; Neurologic Dysfunctions; CNS Demyelinating Autoimmune Disorders; Central Nervous System Autoimmune Demyelinating Diseases; Central Nervous System Autoimmune Demyelinating Disorders; CNS Demyelinating Autoimmune Diseases; Immune destruction