Download Program Overview 11/8/05 - The Research IS Staging Development

The NHLBI Specialized Center of Clinically
Oriented Research (SCCOR) in Pediatric Heart
Disease at the Children's Hospital of
Philadelphia: P50-HL74731
Program Title:
Genetic Mechanisms in Pediatric Heart Disease
http://stokes.chop.edu/programs/sccor/
Program Director: Robert J. Levy, M.D.
The Joseph Stokes, Jr., Research Institute
External Advisory Board
Elazer R. Edelman, MD, PhD, FACC
Director, Harvard-MIT Biomedical Engineering Center
Thomas D. and Virginia W. Cabot Professor
Health Sciences and Technology
Massachusetts Institute of Technology
& Harvard Medical School, Attending Cardiologist,
Brigham and Women’s Hospital
David H. Ledbetter, Ph.D.,
Robert W. Woodruff Professor of Human Genetics
Director, Division of Medical Genetics
Emory University School of Medicine
Jurg Ott, Ph.D.
Professor and Head
Laboratory of Statistical Genetics
Rockefeller University
The Joseph Stokes, Jr., Research Institute
The CHOP SCCOR—Programmatic Hypothesis
“The CHOP SCCOR is a direct outgrowth of productive research
at our Institution over the past decade that was based on the hypothesis that
congenital heart abnormalities are caused by gene defects”
“Basic discoveries concerning gene abnormalities and related patterns of
gene expression can be applied to a unifying approach for both understanding
the complex basis for cardiac dysmorphogenesis as well as providing therapeutic
insights for translational directions.”
The Joseph Stokes, Jr., Research Institute
THE CHOP SCCOR
GENETIC MECHANISMS IN PEDIATRIC HEART DISEASE
PRO JECT
1
2
3
4
5
CORE
A
B
C
D
E
TITLE
Biocompatible Heterograft Biomaterials
Genetic Analysis of Human Outflow Tract M alformations
Genotype and Clinical Outcome in Conotruncal Defects
M olecular Analysis of Human Subtelomeric Rearrangements
Chromosomal Rearrangements and Cardiac Candidate Genes
TITLE
Administrative
Clinical
Cell Culture, DNA and M icroarray
Cardiac M orphology, Gene Expression, and Histology
Bioinformatics and Data Analysis
PROJECT LEADER
Robert Levy, M .D.
Deborah Driscoll, M .D.
Elizabeth Goldmuntz, M .D.
Ian Krantz, M .D.
Beverly Emanuel, Ph.D.
PROJECT LEADER
Robert Levy, M .D.
Elizabeth Goldmuntz, M .D.
Beverly Emanuel, M .D.
Kenneth Ryan, Ph.D.
Peter White, Ph.D.
Academic Unit
Pediatric Cardiology
Genetics
Pediatric Cardiology
Genetics
Genetics
Pediatric Cardiology
Pediatric Cardiology
Genetics
Pediatric Cardiology
Bioinformatics
The Joseph Stokes, Jr., Research Institute
Project 1:Biocompatible Heterograft Biomaterials
Investigations concerning novel surgical therapies for congenital
cardiac malformations.
Project Leader: Robert J. Levy, M.D., Professor of Pediatrics
and Pharmacology, University of Pennsylvania School of
Medicine
Specific Aims
1.
Triglycidyl Amine (TGA)—A new crosslinking reagent for preparing
heart valve bioprostheses: Chemical and biological mechanisms
2. TGA-Matricellular interactions: Cellular and molecular biology studies
related to anticalcification mechanisms.
3. Mechanisms responsible for TGA-mediated inhibition of heart
valve calcification: Biomechanics, biocompatibility, & changes in gene
expression patterns.
Subcontract PI: Joseph Gorman, M.D., Asst.Prof. Surg. Univ.Penn Sch.Med.
Subcontract PI: Michael Sacks, Ph.D., Prof. of Bioengineering, Univ.Pitt.
The Joseph Stokes, Jr., Research Institute
Bioprosthetic Heart Valves
CarpentierHancock
Ionescu-Shiley
Edwards
[Medtronic]
Porcine aortic
valve
Porcine aortic
valve
Bovine pericardium
The Joseph Stokes, Jr., Research Institute
Calcified Bioprosthesis
The Joseph Stokes, Jr., Research Institute
Project 1
Triglycidyl Amine (TGA)
Cl
CH2Cl
O
Epichlorohydrin
HO
OH
N
NH3
i-PrOH-water
Cl
HO
Cl
O
aq. NaOH, toluene-THF
O
N
-HCl
O
TGA
•Reacts irreversibly with lysine, methionine, cystine, histidine
•Results in biomechanical properties superior to glutaraldehyde
•Biocompatibility—supports cellular growth of all cardiovascular cell types
The Joseph Stokes, Jr., Research Institute
Project 2:Genetic Analysis of Human Outflow Tract
Malformations
Project Leader: Deborah Driscoll, M.D., Professor and Chair,
Department of Obstetrics and Gynecology, University of
Pennsylvania School of Medicine
Specific Aims
1.
Developing a map of single nucleotide polymorphisms (SNP’s) in
selected genes implicated in cardiac development.
2. Evaluating SNP’s for potential functional alterations.
3. Determining the genetic contribution of selected genes to the development
of outflow tract malformations using family-based linkage
disequilibrium testing.
4. Identifying modifiers of the cardiac phenotype in patients with 22q11
deletions
The Joseph Stokes, Jr., Research Institute
VEGF Related Directions


Animal models demonstrate VEGF capable of
influencing pharyngeal arch patterning
SNPs with reduced VEGF expression are associated with
cardiovascular defects in 22q11 deletion syndrome
 Identified
an “at risk” haplotype for cardiovascular defects
among individuals with 22q11 deletion

Suggests risk of CHD in the fetus with 22q11 deletion
increases when VEGF levels fall below critical threshold
needed for proper development of pharyngeal arch
arteries, severity may be due to degree of vascular
impairment
The Joseph Stokes, Jr., Research Institute
Project 3:Genotype and Clinical Outcome in Conotruncal
Defects
Project Leader: Elizabeth Goldmuntz, M.D., Associate
Professor of Pediatrics, University of Pennsylvania School of
Medicine
Specific Aims
1.
Investigations of the contribution of NKX2.5 and related genes to the
etiology of controtruncal defects using mutation analyses and familybased association studies.
2. Investigating whether subsets of patients with transposition of the great
arteries or double outlet right ventricle share a common genetic etiology
with the heterotaxy syndrome: Studies of CFC1 mutations & other genes
(NODAL, ZIC3,LEFTY1, ACVRIIB)
3. Studies of the relationship between genetic etiology and clinical
variability/outcome in subjects with conotruncal defects
The Joseph Stokes, Jr., Research Institute
Project 3: Aim 3
Impact of Genotype on Clinical Status
Cross sectional study
 Subjects with TOF, Truncus or IAA
 Ages 8-18 yo
 Clinical Assessment

 Exercise
study
 Echocardiogram
 Cardiac MRI
 Child health questionnaire
The Joseph Stokes, Jr., Research Institute
Project 4:Molecular Analysis of Human Subtelomeric
Rearrangements
Project Leader: Ian Krantz, M.D., Assistant Professor of
Pediatrics and Genetics, University of Pennsylvania School
of Medicine
Specific Aims
1.
Identify individuals with subtelomeric chromosomal deletions and
congenital heart defects.
2. Develop a diagnostic assay that targets the critical region and sizes
subsequent rearrangements of each of the telomeres.
3. Defining the critical regions and identifying candidate disease related
genes for specific clinical phenotypes by mapping the extent and
composition of the associated rearrangements.
The Joseph Stokes, Jr., Research Institute
Project 5:Chromosomal Rearrangements (CR) and Cardiac Candidate Genes
Project Leader: Beverly S. Emanuel, Ph.D., Professor and Chair of
Genetics (at CHOP), University of Pennsylvania School of Medicine
Specific Aims
1. Identify and characterize CR’s in patients with congenital heart disease by high-resolution
cytogenetics and molecular cytogenetic analysis
2. Develop PCR-based mapping strategies using the human genomic sequence to identify the
translocation BP’s
3. Characterize the genomic DNA from normal chromosomes at the chromosomal breakpoints
in order to identify mechanisms of rearrangement
4. Identify the candidate genes disrupted or deleted at the translocation BP’s as candidates for
early cardiac morphogenesis.
5. Determine whether mutations in the candidate genes are associated with the specific cardiac
defect in other patients in the SCCOR Clinical Core.
The Joseph Stokes, Jr., Research Institute
The Clinical Core (a continuing resource from the SCOR’s)
Director: Elizabeth Goldmuntz, M.D.
Objectives:
1. Molecular analyses of the genetic etiology of
conotruncal defects
2. Molecular analyses of bioprostheses
3. Impact of genotype on cardiac anatomy and clinical outcome
Services
1. Ascertain Subjects
2. Acquisition of clinical data
3. Acquisition of relevant samples
4. Review of pertinent medical records
5. Coordination of clinical studies
The Joseph Stokes, Jr., Research Institute
The Cell Culture, DNA, and Microarray Core
(a continuing resource from the SCOR’s)
Director: Beverly Emanuel, Ph.D.
Objectives:
1. Provide cell culture, DNA isolation, cytogenetic and
DNA analysis support for all of the projects
2. Provide microarray resources
3. Training and consultation services to all Projects and
Cores
Services:
1. Establishing lympholastoid cell lines from patients
with congenital heart defects.
2. Isolation of DNA from established cell lines, peripheral lymphocytes
3. Perform FISH to screen for 22q11.2 deletions
4. Regionally localize newly identified human cDNAs by FISH
5. Provide genotyping services for the SCCOR
The Joseph Stokes, Jr., Research Institute
Cardiac Morphology, Gene Expression and Histology Core
Director: Kenneth Ryan, Ph.D., Assistant Professor of Pediatrics,
University of Pennsylvania School of Medicine
Objectives:
1. Analysis of gene (mRNA and protein) expression
2. Histological support
3. Breed mice & xenopus for harvesting embryos for whole-mount
in situ hybridizations and sectioning re. cardiac gene expression patterns
Services
1. Generate and bank frozen staged mouse & xenopus embryo RNA
samples.
2. Generate as blocks and slides embedded mouse and xenopus
embryos.
3. Share expertise in and perform in situ hybridizations using antisense
RNA probes.
4. Perform immunohistochemistry and related histology
The Joseph Stokes, Jr., Research Institute
Bioinformatics and Data Analysis Core
Director: Peter White, Ph.D., Assistant Prof. Ped.,Univ.Penn.School
of Medicine, Director, CHOP’s Bioinformatics Core
Co-Director: Charles Scott, Ph.D.
Objectives:
1. State of the art bioinformatics & biostatistics resources
2. Experimental design support
3. Data base development and data management support
4. The integration of bioinformatics and biostatistics data
on SCCOR subjects
Services
1. Provide assistance in study design, database design,
and data storage
2. To provide infrastructure, hardware, software, technical support,
for analyzing results of molecular biology experiments
3. Statistical analysis and data interpretationThe Joseph Stokes, Jr., Research Institute
NHLBI PEDIATRIC HEART DISEASE
SCCOR’s: 2005 Programmatic Meeting
At the NHLBI, December 13, 2005
Children’s Hospital, Boston
PI: Jane Newburger, M.D.
Children’s Hospital of Cincinnati
PI: Woody Benson, M.D., Ph.D.
Children’s Hospital of Pittsburgh
Steve Webber, M.D., Ph.D.
The Joseph Stokes, Jr., Research Institute
From Molecular Mechanisms to
Improved Outcomes in TOF
Jane W. Newburger, M.D., M.P.H.
Children’s Hospital, Boston
Harvard Medical School
The Joseph Stokes, Jr., Research Institute
Children’s Hospital, Boston:Projects
Project 1: Neurologic and developmental
outcome in TOF (Newburger)
Project 2: Randomized trial of pulmonary valve
replacement in TOF (Geva)
Project 3: Human mutations that cause TOF
(Seidman)
Project 4: Mitochondria in hypertrophied RV and
surgical ischemia (McGowan)
Project 5: Functional analysis of cardiac
transcription factor NKX2.5 (Izumo/Jay)
Project 6: Cardiac regeneration in zebrafish
(Keating)
The Joseph Stokes, Jr., Research Institute
Cores
Core A:
Core B:
Research support and statistics
Microarray core
(Newburger)
(Schinke)
Core C:
Children’s Hospital-Harvard TOF
registry
(Breitbart)
Core D:
Skills development core: The
pathology of CHD
(Collins/Jurazek)
The Joseph Stokes, Jr., Research Institute
Children’s Hospital of Cincinnati: SCCOR in
Pediatric Heart Disease: PI Woody Benson
“Molecular mechanisms of valve development and disease”
PI: D. Woodrow Benson, MD, PhD
Project 1 – Benson—Genetic Studies of Valvular Heart
Disease
Project 2 – Gelb—To Identify PTPN11 Defects Associated
with Noonan’s syndrome and other forms of congenital heart
disease.
Project 3 – Yutzey--Regulation of valvuloseptal
development by DSCR1
Project 4 – Robbins-- Mechanisms of Cardiac pathogenesis
in Noonan Syndrome, effects of SHP-2Gln79Arg, a common
PTPN11 mutation associated with Noonan’s syndrome.
The Joseph Stokes, Jr., Research Institute
Children’s Hospital of Pittsburgh: Optimizing
Outcome after Pediatric Heart Transplantation
SCCOR Program
S.A. Webber, PI
PROJECTS
Consultants
CORES
Project 1
Thymic Tolerance
S. A. Webber, PI
Administration
S. A. Webber, PI
Project 2
Transplant EBV Disease
D. T. Rowe, PI
Immunological
Monitoring
D. Metes, PI
Project 3
Genetic Contributions to
Transplant Outcomes
A. Zeevi, PI
Biostatistics and Data
Management
S. Kelsey, PI
Clinical Skills Training
B. B. Keller, PI
The Joseph Stokes, Jr., Research Institute