An Elite Privilege - Donor Alliance€¦ · 1 C ent er for Hum an Genetics, Duk e Universit y Medic...
Transcript of An Elite Privilege - Donor Alliance€¦ · 1 C ent er for Hum an Genetics, Duk e Universit y Medic...
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An Elite Privilege:
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By Sarah Gray
American Association of Tissue Banks
Director of Marketing and Public Affairs
Why donating for
research is
an incredible honor –
not a consolation prize
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Agenda • The true story of my
infant son’s research donation
• Q & A
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September 30, 2009: First Trimester Screening Results
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Baby A Baby B
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What is Anencephaly? Anencephaly (Greek: without brain) is one of
the most severe forms of a neural tube defect (NTD) and is typically not compatible with life. The neural tube is the developing structure that becomes the brain, skull and spine.
--Duke Center for Human Genetics
The rate of anencephaly in the US is 1 in 4,000 live births. --Center for Disease Control, 2001
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The Birth: March 23, 2010
Inova Fairfax Hospital
Baby A Baby B
Thomas Ethan Gray Callum Mark Gray
10:32 am 10:33 am
4 pounds 1 ounce 5 pounds 10 ounces
APGAR: 4, then 7
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Thomas
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Lived for 6 days
Home death at 1:45 AM,
pronounced by hospice
Transported to hospital by
WRTC van 3:00 AM
Cold solution time 6:53 AM
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Where the donations went
Liver = Cytonet LLC (Durham, NC)
Corneas = Old Dominion Eye Bank (Richmond, VA), then Schepens Eye Research Institute (Boston, MA)
Retinas = University of Pennsylvania (Philadelphia, PA)
Cord Blood = Duke Center for Human Genetics (Durham, NC)
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How I felt about the donation
• He looked perfect at the funeral home.
• Everything happened extremely fast…too fast?
• Did that really just happen?
• There is a person out there somewhere who received my son’s eyes in the mail?
• In other words…
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Grief Counseling at WRTC
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• Met about 20 other
bereaved family
members
• Everyone else donated
to transplant
• Some had already
received letters
• “There, there, research
is important, too.”
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A Conference in Boston: The Accidental Quest Begins
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July 31, 2012 What I saw at Schepens
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Dr. James Zieske
• Senior Scientist, Schepens
Eye Research Institute
• Associate Professor of
Ophthalmology, Harvard
Medical School
Carolyn Bellefeuille
• Annual Giving Officer
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Andrius Kazlauskas, Ph.D.
•Senior Scientist & Sinon
Scholar in Retinal Research,
Schepens Eye Research
Institute
•Professor of Ophthalmology,
Harvard Medical School
Who I met
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Excerpt from email from Dr. James Zieske
August 3, 2012:
“I'm glad that we could reassure you that your donation did make a difference….Your visit helped to remind me that all the eyes we receive are an incredibly generous gift from someone who loved and cared about the person who provided the eyes. I thank you for reminding me of this.”
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November 11, 2012 What we saw at Duke CHG
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Who we met
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• Diedre Krupp, Graduate Student in
Molecular Genetics and Microbiology
• Dr. Simon Gregory, Director, The Duke
Epigenetics and Epigenomics Program
• Allison Ashley-Koch, PhD
Professor of Medical Genetics, Duke
University Medical Center
• Karen Soldano, Lab Research Analyst
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Genetic and Epigenetic Variation in Twins Discordant for Anencephaly
Background
Neural tube defects (NTDs) are a common class of birth defect with complex etiology, thought to be caused by both genetic and environmental factors. Recent studies have implicated epigenetic mechanisms as contributors to NTD causation, esp. DNA methylation (1,2). To investigate how genetic and epigenetic mechanisms contribute to NTDs, we sequenced the exomes and methylomes of a monozygotic twin pair discordant for anencephaly.
Methods
Results: SNV analysis and distribution
In all, 66,231 SNVs were retained after filtering, with an exome-wide Ti/Tv ratio of 2.41. Enrichment for various SNV classes was evaluated against the whole exome for processes previously associated with NTDs, including multiple signaling pathways, one-carbon and folate metabolism, and sphingolipid metabolism. Largely suggestive
Acknowledgements
Samples for this study were collected under NS039818 and funds for the exome and methylation profiling provide by the David H Murdock Research Institute. The authors wish to thank the patients and family members for their participation in and contribution to this NTD study.
Literature Cited
1. Chen et al. Global DNA hypomethylation is associated with NTD-affected pregnancy: a case-control study. Birth Defects Res. A 2010 88:575-581.
2. Chang et al. Tissue-specific distribution of aberrant DNA methylation associated with low-folate status in human neural tube defects. J. Nutr. Biochem. 2011. Epub ahead of print.
• Differential methylation in NTDs is not random: processes relating to neural development are significantly overrepresented among genes with DMRs
• In this twin pair, NTD-associated processes are enriched for intronic and synonymous SNVs, which may have regulatory effects
Intronic SNVs are significantly overrepresented among genes with DMRs
• Findings support potential causative role for altered methylation in NTD etiology, particularly in concert with predisposing genetic variation
Conclusions
Results: Intersection of DMRs and SNVs
455 genes were associated with both DMRs and SNVs. Intronic SNVs were very significantly enriched in these genes (p<2e-16). Intronic DMRs were also enriched (p=0.002). Many GO processes relating to neural development were overrepresented in these genes (Table 3).
P-value Biological Process
1.62E-07 neuron development
2.07E-06 neuron differentiation
3.79E-06 cell morphogenesis involved in neuron differentiation
8.83E-06 neurogenesis
8.89E-06 cell adhesion
5.83E-05 nervous system development
1.29E-04 establishment or maintenance of cell polarity
7.88E-04 face development
1.14E-03 adherens junction
1.61E-03 head development
Deidre Krupp1, Christina A. Markunas1, Karen Soldano1, Kaia S. Quinn1, Heidi Cope1, Melanie E. Garrett1, Allison E. Ashley-Koch1, and Simon G. Gregory1
1Center for Human Genetics, Duke University Medical Center, Durham, NC, USA
Table 3. Selected GO terms overrepresented in genes with both DMRs and SNVs.
P-value Biological Process
2.67E-07 neuron development
1.31E-06 neurogenesis
2.02E-05 nervous system development
3.42E-05 cell morphogenesis involved in neuron differentiation
1.19E-04 cell adhesion
4.06E-04 establishment or maintenance of cell polarity
At the p<1E-05 peak-calling cutoff, 267 genes were associated with hypermethylated (hyper) DMRs in the affected twin, and 503 genes with hypomethylated (hypo) DMRs. 26 genes featured both hypo and hyper DMRs.
Results: DMR analysis and distribution
Genes with hyper DMRs were not enriched for any GO biological processes; genes with hypo DMRs were enriched for multiple NTD-relevant GO processes (Table 2). DNA from blood, fragmented to 200nt
Enrich for coding regions (SureSelect 37Mb kit, Agilent)
Enrich for methylated regions (MethylMiner, Invitrogen)
Sequence 2x150nt (Illumina HiSeq)
Sequence 1x100nt (Illumina HiSeq)
Align to human genome build 37.2 (Burrows-Wheeler Aligner)
Remove unmapped and duplicate reads (Samtools, Picard)
Call SNVs (Genome Analysis ToolKit) Identify DMRs
(Model-based Analysis for ChIP-Seq, significance p<1E-05)
Filter to reduce false positives (read depth>=5,
quality/depth>=10)
Test pathway and DMR enrichment (chi-squared test)
Test for GO term enrichment (g:Profiler, FDR correction,
p<0.05)
Test for SNV enrichment (chi-squared test)
methylome exome
Process P-value SNV type
One-carbon metabolism 0.079
1.04E-06 Synonymous Intronic
Sphingolipid metabolism 0.080 0.070
Exonic Intronic
TGF-b signaling 0.007 Synonymous
WNT signaling (planar cell polarity)
0.025 0.056
Exonic Synonymous
All selected NTD-related terms
0.009 7.28E-09
Synonymous Intronic
enrichments were observed for exonic SNVs, while intronic SNVs were very significantly enriched in several processes (Table 1). These SNVs may have regulatory impact on genes important for neurulation.
Table 1. Selected NTD-related processes with suggestive (0.05 < p < 0.10) or significant (p<0.05) SNV enrichment by chi-squared test.
Table 2. Selected GO terms overrepresented in genes with hypomethylated DMRs.
--Excerpt from Duke Center for Human
Genetics Newsletter, May 2014
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November 11, 2012 What we saw at Cytonet
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Who we met
Mark Johnston, President & COO
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Sonya Meheux, Head of Production/Process Development Jennifer Michaux, Product Development
Scientist
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4/15/10 Outcome letter from Cytonet:
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Excerpt from email from Mark Johnston, Cytonet President & COO:
November 11, 2012:
“I cannot express enough how touched we were by your visit today. I hope you got out of the visit as much as we did. It was a great opportunity for our staff to interact directly with a donor family, as this is something we don’t often get the chance to do.”
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Callum dressing up as a
“doctor” in gown from Cytonet visit
•Specializes in flu shots & vaccines
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August 31, 2013
Old Dominion Eye Foundation
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Who we met: Jennifer Payton, Lab Coordinator
Bill Proctor, CEO, Executive Director
Christina Jenkins, Associate Director – Public Relations Director
Dina Phelps, Technical Director
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Something for Callum’s file
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Meeting NDRI
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NDRI CEO, Bill Leinweber and Jeffrey Thomas, Vice President, Strategic Initiatives
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NRDI List - Example
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Letter from Dr. Arupa Ganguly
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Visit to Penn Lab
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The way I see it… Thomas got into Duke, Penn and Harvard.
He also has a job.
He has co-workers and colleagues
who need him in order to do their jobs.
He is relevant in this community.
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Why apologize?
• “Unfortunately, your loved one’s organs are not suitable for research.
• If you want, we can try
for transplant (or we can
discard.)
• Again, I am so sorry.”
• Why apologize for something awesome, history-making, life-giving, life-saving, life-changing, appreciated and honored?
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Celebrate!
Donating to research is an opportunity to make medical history. It is an opportunity that many of us will never have. Infant corneas and retinas are extremely valuable for
research. This opportunity is a privilege! There is potential to tour the lab.
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Eversight: The Donor Family-Researcher Program
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My wish Share this story with
someone who needs it
Do one thing differently
when it comes to research
donation
Wonder about it out loud with each other
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Upcoming book:
Sept 27, 2016
A LIFE EVERLASTING
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Questions?
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Callum Gray,
modeling his
Harvard and
Duke
t-shirts
Sarah Gray
Director, Marketing & Public Affairs
AATB
703-229-1031