Highthroughput Screening and Cell Repository Core

Director

M. Carrie Miceli, Ph.D.

DMD therapeutic strategies are leading the way in personalized genetic medicine, with several approved and emerging therapies which target only a subset of patients with precisely defined mutations. Core B provides resources to muscle researchers that enable validation of therapeutic strategies and lead compounds drug/chemical libraries, new target identification or basic bio-discovery. We bank and validate human DMD patient derived cells, well defined for DMD mutation, to be used by Center Investigators as well as labs outside the CDMD.

Primary fibroblast, myoblasts, iDRM and iPSC cells will be distributed to individuals requesting them as either frozen vials or expanded live cell stocks. Prior to distribution Core staff will assess the availability of early pass cells, ensuring that some early pass cells remain and that stocks are replenished through expansion. Cells will have been de-identified prior to distribution so recipients are not required to have IRB approval. Distribution of iPSC is more difficult and requires recipient to acquire ESCRO (managed through the UCLA ESCRO office prior to distribution of iPSC). As a result, it is anticipated that the distribution of hiPSC will be more limited. Core B personnel can help facilitate this approval, when appropriate.

We have collected a large number of patient fibroblasts with defined DMD mutations derived from DMD patient skin biopsies. Fibroblasts are first banked, then reprogrammed using two technologies to create iDRM (inducible directly reprogrammed myotubes and iPSC (induced pluripotent stem cells). Initial iPSC were established and characterized as a collaboration with the Broad Stem cell core and Dr. Pyle. Because iDRM are more easily/quickly created and validated, we have created more iDRM than iPSC. The iPSC have the advantage of having potential for differentiation into multiple cell lineages, including skeletal muscle, cardiac and others for studies in which investigators would like to examine on or off target effects of a therapy in different cellular settings.

The patient skin biopsies are directly reprogrammed to iDRM (inducible directly through transduction with a puromycin selectable lent-viral construct expressing tamoxofin inducible MyoD-ER[T]), alone or together a second vector expressing hTERT. Methods of reprogramming iPSC vary, depending on the state of the art at the time they were made. Several more cell lines are being reprogrammed, and our CDMD bank is constantly being expanded. Our bank now contains over 75 DMD, carrier or wild type parent fibroblast samples which are in various stages of reprogramming and development for human models of DMD in a dish. Patient mutations are routinely validated and more finely mapped through Core D (Dr. Nelson, PI).

Most recently we have begun growing and banking primary myoblasts from patient muscle biopsies for use by investigators interested in using them for DMD research studies. Because these myoblasts have not been immortalized and are difficult to expand, they are a limited resource. However, they can be made available to interested investigators on a limited basis and for projects with high priority.

Protocols for best expanding, reprogramming and differentiating DMD cell models are also available through the core. Additional information on patient severity, disease course, access to biopsy tissue, and gene expression is available in some instances through collaboration with Doctors Nelson and Miceli.

To access and updated list of available DMD patient mutations and cell models or for other inquiries, please contact:

M. Carrie Miceli, Ph.D.
Professor of Microbiology, Immunology and Molecular Genetics
Co-Director, Center for Duchenne Muscular Dystrophy
David Geffen School of Medicine and College of Letters and Sciences at UCLA
277B Biomedical Sciences Research Building
cmiceli@ucla.edu

This core is supported by funds from the P30 (Spencer, Miceli and Nelson) and additional grants to Miceli and Nelson from NIH, CIRM, DOD, FED and private philanthropy

Cores

The Core Center is dedicated to understanding processes in degenerative muscle disorders and to identifying therapeutic interventions. In order to most rapidly effect gains, the Center provides a host of interactive intellectual, administrative and research cores. The Center is comprised of four core facilities, all designed to facilitate discovery of therapeutics and test hypotheses related to muscle research. These cores include:

Highthroughput Screening and Cell Repository Muscle Phenotyping and Imaging Bioinformatics and Genomics Back

A primary goal for the Center in creating these Cores is to make available generically useful resources that can be distributed to other researchers in muscular dystrophy or to basic scientists interested in muscle research. The Center is structured to enrich existing muscle research programs on campus by providing its members with access to unique core facilities, pilot funding mechanisms, seminars and an annual retreat focused on muscle research.

Furthermore, the Core Center brings together investigators with a variety of specific interests, some of which are not directly muscle related, to benefit research in muscle disorders.