[R21] The Molecular Basis of R249W LMNA-related Congenital Muscular Dystrophy

Ente: Eunice Kennedy Shriver National Institute of Child Health and Human Development

Scadenza: 2028-05-31

Importo max: 424.743 EUR

Paese: US

Descrizione

ABSTRACT LMNA-related congenital muscular dystrophy (L-CMD) is a rare, devastating genetic disorder that causes severe early-onset muscle weakness and wasting; the R249W mutation accounts for approximately 25% of over 100 reported cases. Despite its prevalence, few studies have investigated R249W in mammals, and no treatments are currently available. This significant knowledge gap hampers understanding of disease mechanisms and the development of therapies. The broad, long-term goal of this project is to understand the molecular basis of R249W L-CMD and explore new therapeutic strategies. We will accomplish this through three specific aims: (1) Examine the effects of the R249W L-CMD mutation on skeletal muscle development; (2) Study how the L-CMD mutation affects lamina organization and the interactions between lamins and their binding partners; and (3) Assess the impact of an adenine base editor (ABE) on the R249W mutation and related phenotypes. The research design employs a multi-faceted approach. For Aim 1, we will characterize R249W’s impact on muscle differentiation. For Aim 2, we will combine computational modeling (AlphaFold, Rosetta) with advanced microscopy techniques (FRAP, Airyscan, TEM) and biochemical methods (immunoprecipitation-mass spectrometry, ChIP-seq) to analyze structural changes in chromatin organization, lamin polymers, and altered protein interactions. For Aim 3, we will design and deliver ABE constructs via lentivirus to patient cells, measure gene correction efficiency (Sanger sequencing, Western blot), and evaluate the rescue of cellular and molecular phenotypes identified in Aims 1 and 2. This project will provide essential mechanistic insights into lamin biology and musculoskeletal development and serve as proof-of-concept for a gene editing therapy. The results will generate vital data and research tools, paving the way for future R01 studies focused on in vivo validation and clinical translation for this overlooked orphan disease. Istituzione: UNIV OF MARYLAND, COLLEGE PARK PI: KAN CAO, Brian G. Pierce Progetto: 1R21HD116032-01A1

Settori: Eunice Kennedy Shriver National Institute of Child Health and Human Development

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