direction of dba research and overview steven r. ellis, ph.d. professor, department of biochemistry...
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Direction of DBA Research and Overview
Steven R. Ellis, Ph.D.Professor, Department of Biochemistry
University of Louisville
July 16th 2015Camp Sunshine
Research Director Diamond Blackfan Anemia Foundation
General Trends in DBA Research
Focus on translational research: Bench to bedside
Continued reliance on private Foundations to support DBA research
• Improved drugs• Genetics; gene discovery, genotype/phenotype relationships
• Beyond anemia; syndromic DBA
• 2012 - 3 year, DOD Award to Dr. Lodish -New drugs for anemia treatment based on a new understanding of the mechanisms of stress erythropoiesis
• April 2014 - bridge grant (DBAF) – between years 1 and 2, $21, 281
• August 2014 – program discontinued
• September 2013 – anticipating funds for year 2
• $95,000 awarded to Dr. Lodish (DBAF and DBAC) to continue his work
Your Foundations at Work
4/2015 – DBAF and DBAC awarded a grant of $35,000 to Johan Flygare to help start up a new lab and screen a library of over 12,000 chemical compounds to identify potential drugs that alleviate the symptoms of their mouse model of DBA
• 2008/2009 Johan was a post-doctoral fellow in Harvey Lodish’s laboratory and did the initial work on how glucocorticoids stimulate red cell production
• Prior to 2008, Johan was a graduate student in the laboratory of Stefan Karlsson helping develop the mouse model for DBA which is being used to test gene therapy approaches for DBA
4/2014 – DBAF awarded a grant of $36,000 to Vijay Sankaran to study the role of GATA1 in red cell production with an eye toward novel therapies for DBA
1/2015 – DBAF and DBA UK awarded a grant of $41,810 to Nickolas Watkins to study the role of RPL5 and RPL11 in signaling p53 activation
Physicians li
ke Drs. Vlachos a
nd Lipton will
have their h
ands full w
ith cli
nical tr
ialsIncreased number of potential drugs
coming down the R&D pipeline
Drugs specifically directed at DBA /drugs targeted at the intrinsic nature of genetic diseases
Personalized medicine, drugs as diverse as the heterogeneity of DBA patients
Direction of DBA Research (2015 Predictions)
• A drug may target a subset of DBA genes
• A drug may target a particular type of genetic lesion
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
Central Dogma of Molecular Biology
mRNA
Protein
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
aagcuuccaau//gguccaucguac
transcription
splicing
pre-mRNA
translation
3.5 billion bases
intronexon
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
mRNA
Protein
translationC
Phe
silent/synonymous
U
Term
nonsense/nonsynonymous
frameshift
Mutation Muddle
G
Glu
missense/nonsynonymous
Lys NH3+ Glu COO -
Asp COO -
*
With the Advent of Whole Exome and Whole Genome Sequencing We are Faced with an Increasing Number of
Variants of Unknown Significance (VUS)
An extended family seen by the bone marrow failure unit at the National Cancer Institute• 3 affected family members with a VUS in RPS19• V138L, valine to leucine substitution at position 138
COO -
C HH3+ N
CH
CH3 CH3
COO -
C HH3+ N
CH
CH3 CH3
CH2
V, Valine L, Leucine
LVVIL
human
Gregory (2007) NAR
• The in silico prediction programs are not unanimous about whether this would be a potentially disease causing variant. It does not exist in any public databases.
V138L
The Plot Thickens
Patients Display a Pre-rRNA Processing Signature Consistent with V138L Being a Pathogenic Mutation
Flygare, Aspesi et al. 2007 Blood
RPS19 Knockdown
✔
✔
PP C
✔
FINAL THOUGHTS
These Folks Represent Only a Fraction of the People Out There Working to Help Your Kids
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
mRNA
Protein
translationC
Phe
silent/synonymous
U
Term
nonsense/nonsynonymous
frameshift
Mutation Muddle
G
Glu
missense/nonsynonymous
Atalurin – a drug that causes ribosomes to read through premature termination codons
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
Mutation Muddle
mRNA
Protein
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
aagcuuccaau//gguccaucguac
transcription
splicing
pre-mRNA
translation
Drugs are in development to fix splicing defects
Physicians li
ke Drs. Lip
ton and Vlachos will
have their h
ands full w
ith cli
nical tr
ialsIncreased number of potential drugs
coming down the R&D pipeline
Drugs specifically directed at DBA /drugs targeted at the intrinsic nature of genetic diseases
Personalized medicine, drugs as diverse as the heterogeneity of DBA patients
Specific Trends in DBA Research
• A drug may target a subset of DBA genes
• A drug may target a particular type of genetic lesion