Transcript Ellis - Diamond Blackfan Anemia Foundation, Inc.

Direction of DBA Research and Overview
July 16th 2015
Camp Sunshine
Steven R. Ellis, Ph.D.
Professor, Department of Biochemistry
University of Louisville
Research Director
Diamond Blackfan Anemia Foundation
General Trends in DBA Research
Focus on translational research: Bench to bedside
• Improved drugs
• Genetics; gene discovery, genotype/phenotype relationships
• Beyond anemia; syndromic DBA
Continued reliance on private Foundations to support DBA research
• 2012 - 3 year, DOD Award to Dr. Lodish -New drugs for anemia
treatment based on a new understanding of the mechanisms of stress
erythropoiesis
• September 2013 – anticipating funds for year 2
• April 2014 - bridge grant (DBAF) – between years 1 and 2, $21, 281
• August 2014 – program discontinued
• $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
Direction of DBA Research (2015 Predictions)
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
• A drug may target a
subset of DBA genes
• A drug may target a
particular type of genetic
lesion
Central Dogma of Molecular Biology
transcription
pre-mRNA
aagcuuccaau//gguccaucguac
3.5 billion bases
intron
exon
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
mRNA
splicing
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
translation
Protein
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
Mutation Muddle
mRNA
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
frameshift
C
U
G
translation
Protein
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
Phe
silent/synonymous
Term
Glu
nonsense/nonsynonymous
Lys NH3+
Asp COO -
Glu COO -
missense/nonsynonymous
*
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 H3+ N
C
H
CH
CH3
CH3
COO H3+ N
C
CH2
CH
CH3
V, Valine
H
CH3
L, Leucine
The Plot Thickens
• 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
L
V
V
I
L
human
Gregory (2007) NAR
Pre-rRNA Processing as a Means of Studying
Ribosomal Protein Function
Patients Display a Pre-rRNA Processing Signature
Consistent with V138L Being a Pathogenic Mutation
P C P
RPS19
Knockdown
✔
✔
✔
Flygare, Aspesi et al. 2007 Blood
FINAL THOUGHTS
These Folks Represent Only a Fraction of the People
Out There Working to Help Your Kids
Mutation Muddle
mRNA
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
frameshift
C
U
G
translation
Protein
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
Phe
silent/synonymous
Term
Glu
nonsense/nonsynonymous
missense/nonsynonymous
Atalurin – a drug that causes
ribosomes to read through
premature termination codons
Mutation Muddle
transcription
pre-mRNA
aagcuuccaau//gguccaucguac
Drugs are in development to fix
splicing defects
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
mRNA
splicing
AUG – UUU – AGG – CUG – GCA – AAG – CUA – AAA – GGA – UCC - UAA
translation
Protein
Met – Phe – Arg – Leu – Ala – Lys – Leu – Lys – Gly – Ser - Term
Specific Trends in DBA Research
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
• A drug may target a
subset of DBA genes
• A drug may target a
particular type of genetic
lesion