Charles P. Emerson, Jr. - US grants

The Cell Core C is the keystone of the FSHD-Wellstone research activities and will be responsible for the development and maintenance of reagents for its investigators and the FSHD research community at-large. Every project in this proposal will draw from its resources. A FSHD tissue bank (Aim 1) will be established from patients and first-degree family members specifically recruited, screened and enrolled to provide high quality and highly relevant biological specimens. Open muscle biopsies of FSHD muscle and control muscle will be obtained from 3 participating sites: Johns Hopkins, University of Utah and University of Sao Paulo, and distributed to multiple institutions for studies proposed. The Cell and Molecular Lab (Aim 2) of the Core will be located at BBRI and will perform several functions including the establishment of primary and secondary cultures and selected immortalized clonal lines from biopsy materials of FSHD subjects and primary relatives. Additionally, evaluation of biochemical, cell biological, proteomic and molecular data obtained from the relevant cell cultures and immortalized cell lines will be made available to the FSHD scientific community through the establishment of a national resource FSHD muscle cell repository. An FSHD myoblast cell repository (Aim 3) will be established through collaboration with Genzyme/Myosix and an FSHD lymphocyte repository will be established with The Coriell Institute. Finally the Bioinformatics Support Group (Aim 4) will have a large role in this Center, serving all projects as well as individual aspects of the Cell Core. The Bioinformatics Support Group will set-up and maintain a portal for standardizing, managing and sharing data between collaborating laboratories in the Center, and for making data publicly available as a companion to the public myoblast cell repository. It will also perform the computational analysis and modeling of the data produced by Cell Core C, including transcriptomic data, proteomics data, and data from cell-based assays. The Bioinformatics component will use both statistical and machinelearning approaches to identify FSHD biomarkers from across the different data modalities, and will perform integrative analysis, with the goals of understanding the pathways that are disrupted in FSHD and identifying and assessing potential therapeutic targets.

Facioscapulohumeral muscular dystrophy (FSHD) is a complex genetic disorder resulting in the progressive and selective degeneration of the FSH muscles in adults. Many histopathological features of FSHD mirror muscle aging and loss of regenerative capacity, as evidenced by reduced populations of myogenic progenitors in affected muscles and their defective growth and survival behavior in cell culture. The disease is linked to the contraction of D4Z4 repeats in the subtelomeric region of Chromosome 4, leading to the idea that deletion of these repeats results in the overexpression of adjacently located genes or to Telomere Position effects that lead to telomere shortening and cellular aging. The central hypothesis to be tested in Project 3 is that FSHD is a regenerative disease caused by genetic regulatory disruptions that reduce the viability and/or telomere function in myogenic progenitors, resulting in premature cell aging and consequent loss of muscle regenerative capacity. Immunohistological, cell culture and biomarker approaches will be used to systematically investigate myogenesis in affected and unaffected muscles recovered by surgical biopsy of a large cohort of FSHD patients and first-degree relatives. The goals of these studies are: 1) to identify biomarkers expressed in FSHD muscle and muscle progenitors for monitoring the efficacy of FSHD clinical trials;2) to identify disease-specific biomarkers that have functional roles in FSHD muscle pathology as targets for development of drugs, RNAi, and cell-based therapeutics that promote satellite cell survival, myogenesis and improved muscle regeneration;and 3) to establish validated cell and mouse models of FSHD. Biomarker and telomere studies will enable investigations of the underlying molecular pathology of FSHD to test D4Z4 locus control and telomere position effect (TPE) disease mechanisms. Project 3 benefits from the distinct multidisciplinary expertise of Center investigators and collaborators working in an interactive research environment and from the exceptional resources and technical support of the Cell Core with its unique collection of FSHD muscle biopsies and muscle cell cultures. The proposed studies will elucidate the disease role of myogenesis and muscle regeneration in FSHD and provide validated cell models for investigations of FSHD disease mechanisms and the development of drug and cell-based therapeutics for the treatment of FSHD, a currently untreatable disease.

A-152E5.1; ABCD-1; Adam11; Adam11 gene; Advisory Committees; Advocate; Area; Bio-Informatics; Biocompatible Materials; Bioinformatics; Biomaterials; Body Tissues; Businesses; CCL22; CCL22 gene; CLB/DACT/MTX; Cells; Chlorambucil/Dactinomycin/Methotrexate; Clinical; Clinical Management; Collaborations; Communication; Communities; Computer Programs; Computer software; Congresses; Contracting Opportunities; Contracts; Core Facility; DC/B-CK; Data; Development; Direct Costs; Education; Educational aspects; Environment; Evaluation; Event; Facioscapulohumeral Atrophy; Facioscapulohumeral Muscular Dystrophy; Funding; Goals; Human Resources; Industry; Information Technology; Institution; Internet; Investigators; Landouzy Dejerine muscular dystrophy; Landouzy-Dejerine Dystrophy; Leg; MDC; MDC protocol; MGC34554; Mammals, Mice; Manpower; Mice; Mission; Murine; Mus; Muscular Dystrophies; Muscular Dystrophy, Facioscapulohumeral; Muscular Dystrophy, Fasioscapulohumeral; Myodystrophica; Myodystrophy; NIH; National Institutes of Health; National Institutes of Health (U.S.); P01 Mechanism; P01 Program; Participant; Patient Education; Patient Instruction; Patient Representative; Patient Training; Patients; Persons; Policies; Postdoc; Postdoctoral Fellow; Procedures; Program Project Grant; Program Research Project Grants; Programs (PT); Programs [Publication Type]; Progressive Muscular Dystrophy, Facioscapulohumeral Type; Publications; Records; Reporting; Research; Research Activity; Research Associate; Research Personnel; Research Program Projects; Research Resources; Research Training; Researchers; Resources; SCHED; SCYA22; STCP-1; Schedule; Scientific Publication; Scientist; Services; Site; Software; Task Forces; Tissues; Training Programs; Training and Education; Travel; United States National Institutes of Health; WWW; Writing; chlorambucil/dactinomycin/methotrexate protocol; computer program/software; conference; cost; day; experience; interest; member; outreach; personnel; post-doc; post-doctoral; programs; repository; symposium; web; wiki; world wide web

DESCRIPTION (provided by applicant): Hedgehog (Hh) signaling plays an important role in embryonic patterning and adult stem cell renewal, but has recently been found also to be involved in certain stem-cell cancers, including pancreatic, small cell lung, intestinal, and prostate, all of which are notoriously difficult to treat. One of the first steps in Hh signaling is the autoprocessing of Hh protein, in which the C-terminal domain (Hh-C) catalyzes a cholesterol-dependent autocleavage reaction that leads to the production of the cholesterol ester of the N-terminal Hh domain (Hh-N), thereby yielding a signaling molecule. The secreted Hh ligand binds to the Patched receptor, leading to activation of Smoothened (Smo) and a signal transduction cascade that culminates in the activation of Gli transcription factors and the transcription of Gli and other genes. Most research on the role of Hh signaling in stem cell cancers is focused on the series of events that lie between the activation of Smo and the activation of Gli, with the aim of understanding this complex signaling pathway and in the hope to develop cancer therapeutics. In contrast, very little attention has been given to the role of Hh autoprocessing and modification, in spite of the fact that it represents the initial step in the Hh signaling pathway. Due to the absence of pharmacologic tools for its study, there is considerable uncertainty about the role of Hh autocleavage and esterification with cholesterol in Hh signaling, both in Hh signaling in the developing embryo and in the growth of Hh ligand-dependent cancers. This proposal aims to identify compounds that attenuate Hh autoprocessing as a research tool for dissecting the role of autoprocessing and the attendant cholesterol modification in Hh function, both in the many different contexts of embryonic development and in Hh ligand-dependent stem cell cancers. This approach is based on an in vitro homogeneous assay system which measures changes in fluorescence polarization that accompany the cholesterol-depended autocleavage of Hh protein. Of special interest will be the study of the effect of the inhibitors identified by this research on the growth of Hh-dependent endodermal tumor cell lines. If growth inhibition of the tumor cell lines should indeed be observed, this would suggest that Hh autoprocessing, as the first step in the Hh signaling pathway, could be a possible therapeutic target. PUBLIC HEALTH RELEVANCE: Hedgehog (Hh) signaling plays an important role in embryonic patterning and adult stem cell renewal, but has recently been found also to be responsible for certain stem-cell cancers, including pancreatic, small cell lung, intestinal, and prostate, all of which are notoriously difficult to treat. The first step in Hh signaling is an autoprocessing reaction, but there are no chemical biology tools available for studying this important step. This proposal seeks to identify inhibitors of Hh autoprocessing that can serve as tools for studying its role in normal development and in cancer and which may lead to potential cancer therapeutics.