Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset neurological disease in which motor neurons degenerate, thereby leading to paralysis and death due to respiratory failure. The incidence of ALS differs based on ancestral origin, with ~5.5 cases per 100, 000 persons in the United States. In addition to motor function impairment, a subset of patients (~15%) suffers from frontotemporal dementia (FTD), leading to low quality of life and extreme emotional strain on the patient's caregivers. From a genetic perspective, ALS pathogenesis is commonly driven by GGGGCC (G4C2) repeat expansions in the C9ORF72 gene, which cause a disease subtype of ALS referred to as c9ALS/FTD. The repeat expansions lead to C9ORF72 haploinsufficiency, accumulation of repeat RNA, and the production of aggregation-prone proteins composed of repeating dipeptides. While antisense oligonucleotides and RNA interference (RNAi) approaches have been used to target the causal mutations of ALS, these approaches are limited due to their transient effect, lack of specificity, and inability to multiplex. Gene editing technologies have emerged as powerful approaches to target causal drivers of rare diseases and engineer cell-based therapeutics. Therefore, there is a critical need for a gene editing-based therapy that can safely and efficiently deplete the disease-causing mutant C9ORF72. The original CRISPR/Cas9 system that targets DNA is limited in this context since repeat expansions are at the kilobase scale and cannot be efficiently deleted with current Cas9-based technologies. The overall goal of this proposal is to develop a gene therapy for ALS using the Acrobat Genomics proprietary enARGN gene editing system, which is more compact than the CRISPR/Cas9 system and more deliverable. Preliminary studies have demonstrated the activity of Acrobat Genomics ARGN gene editing system. In this Phase I SBIR project, Acrobat Genomics plans to develop a high-throughput protein engineering platform (AcrobaTx) to enhance ARGN gene editing activity. The protein engineering approach is enabled by advancements in high-throughput oligonucleotide synthesis, sequencing technologies, and protein language models. Notably, the approach utilizes pooled screening and is highly parallel to enable the study of thousands of proteins in a single experiment (Specific Aim 1). With an optimized ARGN protein, a novel editing strategy will be developed to deplete expression of mutant C9ORF72 and restore expression of the wild-type allele (Specific Aim 2). In summary, by protein engineering the ARGN system, Acrobat Genomics will develop a new therapy for ALS, which is currently a devastating and incurable disease with few treatment options. In addition to the proposed research investigations, this application will lay the foundation for the entrepreneurial career development of the PI. The PI is the co-inventor of foundational technologies used in this proposal and has co-founded Acrobat Genomics. The PI aims to transition from academia to commercialize the enARGN system as a therapeutic for genetic diseases and has identified mentors with significant business and scientific expertise to aid in this endeavor.
Public Health Relevance Statement: Narrative Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset neurological disease in which motor neurons degenerate, thereby leading to paralysis and death due to respiratory failure. There are no existing therapies that specifically target the repeat expansions in the C9ORF72 gene that are known to cause ALS. Acrobat Genomics will protein engineer optimized and compact RNA- guided nucleases to precisely target and deplete repeat-bearing C9ORF72 transcripts as a potential therapeutic for patients with ALS. Terms: <2-Amino-6-trifluoromethoxybenzothiazole; 21+ years old; 65 and older; 65 or older; 65 years of age and older; 65 years of age or more; 65 years of age or older; 65+ years; 65+ years old; > 65 years; ALS therapy; ALS treatment; Academia; Address; Adult; Adult Human; Adverse Experience; Adverse event; Age of Onset; Aged 65 and Over; Airway failure; Alleles; Allelomorphs; Amyotrophic Lateral Sclerosis; Amyotrophic Lateral Sclerosis Motor Neuron Disease; Antisense Agent; Antisense Oligonucleotides; Assay; Bioassay; Biological Assay; Biotech; Biotechnology; Businesses; C9ALS/FTD; C9ORF72; CRISPR approach; CRISPR based approach; CRISPR method; CRISPR methodology; CRISPR technique; CRISPR technology; CRISPR tools; CRISPR-CAS-9; CRISPR-based method; CRISPR-based technique; CRISPR-based technology; CRISPR-based tool; CRISPR/CAS approach; CRISPR/Cas method; CRISPR/Cas technology; CRISPR/Cas9; CRISPR/Cas9 technology; Capital; Care Givers; Caregivers; Cas nuclease technology; Cell Body; Cell Therapy; Cells; Cessation of life; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats approach; Clustered Regularly Interspaced Short Palindromic Repeats method; Clustered Regularly Interspaced Short Palindromic Repeats methodology; Clustered Regularly Interspaced Short Palindromic Repeats technique; Clustered Regularly Interspaced Short Palindromic Repeats technology; Complementary DNA; Couples; DNA; DNA Therapy; Death; Deoxyribonucleic Acid; Development; Dipeptides; Disease; Disorder; Doctor of Philosophy; Down-Regulation; Dysfunction; Emotional; Engineering; Event; FDA approved; Foundations; Frontal Temporal Dementia; Frontotemporal Dementia; Functional disorder; Gehrig's Disease; Gene Transfer Clinical; Generations; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Diseases; Genetic Intervention; Genetic defect; Genomics; Goals; Guide RNA; High Throughput Assay; Human; Human Cell Line; Impairment; Incidence; Investigation; Investigators; Knock-in; Knock-out; Knockout; Language; Lead; Link; Lou Gehrig Disease; Machine Learning; Measurement; Measures; Mentors; Mitochondria; Modeling; Modern Man; Motor; Motor Cell; Motor Neurons; Mutation; Names; Nervous System Diseases; Nervous System Disorder; Neurologic Disorders; Neurological Disorders; Non-Polyadenylated RNA; Oligo; Oligonucleotides; Orphan Disease; Palsy; Paralysed; Pathogenesis; Pathogenicity; Patients; Pb element; Peptides; Persons; Ph.D.; PhD; Phase; Phase 3 Clinical Trials; Phase III Clinical Trials; Physiopathology; Plegia; Post-Transcriptional Gene Silencing; Posttranscriptional Gene Silencing; Production; Protein Engineering; Proteins; QOL; Quality of life; RNA; RNA Gene Products; RNA Interference; RNA Silencing; RNA Splicing; RNA-Binding Proteins; RNAi; Rare Diseases; Rare Disorder; Research; Research Personnel; Researchers; Respiratory Failure; Ribonucleic Acid; Riluzole; SBIR; SOD-1; SOD-1 protein; SOD1; SOD1 gene; SOD1 gene product; Sequence-Specific Posttranscriptional Gene Silencing; Small Business Innovation Research; Small Business Innovation Research Grant; Specificity; Splicing; System; TAR DNA-binding protein 43; TDP-43; TDP43; Technology; Therapeutic; Toxic effect; Toxicities; Transcript; United States; Universities; Variant; Variation; Work; above age 65; adulthood; after age 65; age 65 and greater; age 65 and older; age 65 or older; age > 65; age of 65 years onward; aged 65 and greater; aged 65+; aged â¥65; amyotrophic lateral sclerosis therapy; amyotrophic lateral sclerosis treatment; antisense oligo; base editing; c9FTD/ALS; c9als/frontotemporal dementia; c9ftd/amyotrophic lateral sclerosis; cDNA; career development; causal allele; causal gene; causal mutation; causal variant; causative mutation; causative variant; cell mediated therapies; cell-based therapeutic; cell-based therapy; cellular therapeutic; cellular therapy; chromosome 9 open reading frame 72; commercialization; developmental; disease subgroups; disease subtype; disorder subtype; edarabone; edaravone; experiment; experimental research; experimental study; experiments; front temporal dementia; frontal lobe dementia; frontotemporal lobar dementia; frontotemporal lobe degeneration associated with dementia; gRNA; gene editing platform; gene editing system; gene editing technology; gene editing tools; gene repair therapy; gene therapy; gene-based therapy; gene-editing toolkit; genetic condition; genetic disorder; genetic protein engineering; genetic therapy; genome mutation; genomic therapy; heavy metal Pb; heavy metal lead; high throughput screening; human old age (65+); iPS; iPSC; iPSCs; in vivo; induced pluripotent cell; induced pluripotent stem cell; inducible pluripotent stem cell; knockin; loss of function; machine based learning; mitochondrial; motoneuron; motor neuron degeneration; mutant; name; named; naming; neurological disease; neuron toxicity; neuronal toxicity; neurotoxicity; new drug treatments; new drugs; new pharmacological therapeutic; new therapeutics; new therapy; next generation therapeutics; norantipyrine; norphenazone; novel; novel drug treatments; novel drugs; novel pharmaco-therapeutic; novel pharmacological therapeutic; novel therapeutics; novel therapy; nuclease; old age; older adult; older adulthood; oligos; orphan disorder; over 65 years; paralysis; paralytic; pathophysiology; patient population; patient subclass; patient subcluster; patient subgroups; patient subpopulations; patient subsets; patient subtypes; phase III protocol; phenylmethylpyrazolone; protein TDP-43; protein TDP43; protein design; screening; screenings; superoxide dismutase 1; therapeutic agent development; therapeutic development; timeline; vector; â¥65 years
