Gene therapy provides a functional cure for patients with genetic disorders and is particularly relevant to the treatment of monogenic rare diseases (those caused by a single gene), such as Duchenne muscular dystrophy (DMD). Adeno-associated virus (AAV) is the most prevalent and clinically successful viral gene therapy. However, as a delivery platform, it exhibits significant limitations when it comes to immunogenicity, cargo ca- pacity, manufacturability, and ease of cell type-specific targeting. Other platforms, such as lipid nanoparti- cle/mRNA formulations, can be manufactured at scale but only induce untargeted and transient gene expres- sion. Therefore, while each has its strengths and weaknesses, there is currently no platform available that brings accessible and durable gene therapy to rare genetic disorders. The goal of M13-Tx is to develop a next-generation in vivo gene delivery platform that addresses the current key limitations of existing gene therapy technologies. This platform is based on phage-derived particles (PDPs) - an immune-privileged, easily engineerable, and efficiently produced derivation of M13 bacteriophage - spe- cifically designed for the transduction of human cells. By the combination of directed evolution and rational de- sign, these PDPs can be engineered to target any tissue and deliver up to 20kb DNA cargo. In this Phase I SBIR, M13-Tx proposes to engineer and screen PDPs to successfully deploy a cargo carrying the functional copy of DMD gene coding for dystrophin protein in muscle tissue in vivo. In contrast to existing AAVs, which can only pack a fraction of the DMD coding sequence, the PDPs will deploy the full DMD gene only in muscle tissue for durable and redoseable treatment of Duchenne muscular dystrophy. The final product of this proposal is lead PDPs, ready to be tested in the DMD mouse model. The lead PDPs and muscle-specific deployment can be utilized across rare diseases caused by mutations of different genes but manifested in the same tissue (e.g., Myotonic Dystrophy and Facioscapulohumeral Muscular Dystrophy). Moreover, the same workflow can be used to find PDPs for other difficult-to-target tissues (e.g., lung and CNS). The proposed study will serve as a proof-of-concept, showing that PDPs can be selected for specific gene delivery purpose, given their engineerable coat and easily modified cargos. Developing this technology will revolutionize gene therapy by presenting a versatile and affordable gene delivery platform.
Public Health Relevance Statement: PROJECT NARRATIVE There is currently no affordable and redoseable gene therapy that can deliver a large DNA cargo to specific tissues and trigger durable correction of the underlying genetic problem while evading immune surveillance. M13-Tx proposes to engineer a next-generation in vivo gene delivery technology based on rational design, screening and in vivo selection of phage-derived particles (PDP), capable of delivering a full dystrophin gene to muscle cells for the treatment of Duchenne muscular dystrophy (DMD). Both the PDP delivery and deployment platform, and the workflow for selecting the best-performing in vivo variants, are generalizable across rare dis- eases.
Project Terms: Adeno-Associated Viruses; Dependoparvovirus; adeno associated virus group; Dependovirus; Affect; Bacteriophages; Phages; bacterial virus; Bar Codes; barcode; Biomedical Engineering; bio-engineered; bio-engineers; bioengineering; biological engineering; Capsid; Cell Nucleus; Nucleus; Cells; Cell Body; Cytomegalovirus; CMV; HCMV; Salivary Gland Viruses; cytomegalovirus group; DNA; Deoxyribonucleic Acid; Duchenne muscular dystrophy; Duchene; Duchenne; Duchenne-Griesinger syndrome; Ellis-van Creveld (EvC) syndrome; Pseudohypertrophic Muscular Dystrophy; X-linked dilated cardiomyopathy; X-linked muscular dystrophy; X-linked recessive muscular dystrophy; benign X-linked recessive muscular dystrophy; childhood pseudohypertrophic muscular dystrophy; classic X-linked recessive muscular dystrophy; mild X-linked recessive muscular dystrophy; progressive muscular dystrophy of childhood; pseudohypertrophic adult muscular dystrophy; pseudohypertrophic muscular paralysis; Engineering; Evolution; Exhibits; Future; Gene Expression; gene therapy; DNA Therapy; Gene Transfer Clinical; Genetic Intervention; gene repair therapy; gene-based therapy; genetic therapy; genomic therapy; Genes; Viral Genes; Goals; Human; Modern Man; Immunity; Immunologic Surveillance; Immune Surveillance; Immunologic Surveillances; Immunological Surveillance; Immunological Surveillances; Immunosurveillance; In Vitro; Lead; Pb element; heavy metal Pb; heavy metal lead; Libraries; Lipids; Liver; hepatic body system; hepatic organ system; Lung; Lung Respiratory System; pulmonary; Muscle; Muscle Tissue; muscular; Muscular Atrophy; Muscle Atrophy; muscle breakdown; muscle degradation; muscle deterioration; muscle loss; muscle wasting; Myopathy; Muscle Disease; Muscle Disorders; Muscular Diseases; Myopathic Conditions; Myopathic Diseases and Syndromes; Myopathic disease or syndrome; muscular disorder; Myotonic Dystrophy; Dystrophia Myotonica; Myotonia Atrophica; Myotonia Dystrophica; Steinert Disease; dystrophic myotonia; Patients; Proteins; Resources; Research Resources; Retina; RNA; Non-Polyadenylated RNA; RNA Gene Products; Ribonucleic Acid; Messenger RNA; mRNA; Technology; Testing; Tissues; Body Tissues; Transfection; Translations; translation; Work; Dystrophin; promotor; promoter; Coliphage M13; Enterobacteria phage M13; M13 Phage; Bacteriophage M13; improved; Surface; Clinical; Phase; Variation; Variant; Muscle function; muscle degeneration; FSHD; Facioscapulohumeral Atrophy; Facioscapulohumeral Type Progressive Muscular Dystrophy; Fasioscapulohumeral Muscular Dystrophy; Landouzy Dejerine muscular dystrophy; Landouzy-Dejerine Dystrophy; Facioscapulohumeral Muscular Dystrophy; directed evolution; Directed Molecular Evolution; Therapeutic; Genetic; Deposition; Deposit; Immune; Immunes; Clinic; cell type; Nuclear; Muscle Cells; Myocytes; Myoblasts; Embryonic Muscle Cells; Precursor Muscle Cells; particle; trafficking; Rare Diseases; Orphan Disease; Rare Disorder; orphan disorder; Code; Coding System; design and construction; design and construct; Muscle Development; Muscular Development; Coat Proteins; Viral Coat Proteins; Viral Outer Coat Protein; Capsid Proteins; protein expression; Address; Data; Mammalian Cell; in vivo; Genetic Medicine; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Gene Delivery; Derivation procedure; Derivation; Development; developmental; Output; immunogenicity; designing; design; next generation; Biodistribution; transduction efficiency; muscular dystrophy mouse model; disease-causing mutation; effective treatment; effective therapy; full scale manufacturing; large scale manufacturing; mass production; large scale production; screenings; screening; targeted drug therapy; targeted drug treatments; targeted therapeutic; targeted therapeutic agents; targeted therapy; targeted treatment; Formulation; immune evasive; Immune Evasion; genetic condition; genetic disorder; Genetic Diseases; manufacturability; rare genetic disease; rare genetic disorder; delivery vehicle; delivery vector; rational design; manufacture; manufacturing systems
