Transcript Genetics of Rheumatoid Arthritis in some Arab States

Genetics of Rheumatoid Arthritis
in some Arab States
Thurayya Arayssi M.D
Weill Cornell Medical College-Qatar
May 2012
Welcome
Objectives of the Meeting
• To review study design
• To discuss study details
• To familiarize investigators with study
procedure
• To assess the needs of the collaborating
centers for a successful study
Aims of the Study
1. Enroll and Collect DNA on at least 2000 RA
cases and 2000 controls
2. Assess for the defined RA risk alleles by
genotyping a subset of 500 seropositive cases
and 500 controls
3. Sequence the whole exome in a large
consanguineous family to discover inherited
recessive protein coding mutations that
cause RA.
Common Alleles Associated with RA
Risk
Figure 2: Common alleles associated with risk of RA. Timeline of discovery of common
alleles from candidate gene and genome-wide studies. These RA risk alleles have not yet
been tested among individuals of Arab ancestry.
T-Cell Activation
NF Kappa B pathway
HLA DRB1:involved in MHC molecule–based antigen
REL:Proto-oncogene member of the NF-κB family that
presentation and responsible for self-peptide selection and Tcell repertoire
regulates leukocyte activation and survival
PTPN22:Lymphocyte-specific nonreceptor tyrosine
TNFAIP3:Signaling protein and negative regulator of TNF-α–
phosphatase involved in regulation of activation threshold of
lymphocyte
induced NF-κB activation
AFF3:Transcription factor for lymphoid development
TRAF1:Regulator of TNF-α–receptor superfamily signaling
(e.g., to NF-κB and JNK)
CD28:Costimulatory molecule for T-cell activation
Other pathways
CD40:Costimulatory molecule that enhances interactions
BLK:B-lymphoid tyrosine kinase involved in B-cell receptor
between T and B cells and increases auto- antibody production
signaling and B-cell development
CTLA4:Costimulation suppressor that regulates interactions
CCL21
between T cells and antigen-presenting cells
IL2RA:High-affinity receptor for interleukin-2 on lymphocyte
FCGR2A
subsets
IL2:Cytokine that regulates activation of T cells, particularly
PADI4:Enzyme that converts arginine to citrulline, creating
regulatory T cells
autoantigens in rheumatoid arthritis
IL-21:Cytokine that regulates differentiation of T cells,
PRDM1
particularly Th17, and activation of B cells
PRKCQ, STAT4, TAGAP:Member of the protein kinase C family
that regulates T-cell and macrophage activation; Transducer of
cytokine signals that regulate proliferation, survival, and
differentiation of lymphocytes; Rho-GTPase enzyme involved in
T-cell activation
NEJM, 365;23nejm.orgdecember 8, 2011
TNFRSF14
Studies from the Middle East
• Largest study by Dr. Kazkaz:
– A dose effect was observed between the SE copy
number and
• risk of RA (OR 1 vs 0 copies = 1.6; OR 2 vs 0 = 15.3)
• radiographic joint destruction (OR 1 vs 0 = 2.2; OR 2 vs
0 = 9.9)
Our study
• Genome Wide Association Study (GWAS) that
will test for >30 identified risk alleles
identified in European and North American
populations
Patients
Inclusion Criteria
•
•
•
•
•
Self reported Arab ancestry
Age > 18
RA diagnosed per the ACR criteria
Ability to provide informed consent
Otherwise no exclusion criteria
Case log
Controls
Inclusion Criteria
• Self reported Arab Ancestry
• Absence of an underlying autoimmune
disorder
• Able to provide an informed consent
• 1:1 matching for age ( 10 year range), gender
and reported ancestry
Control log
Projected number of patients/center
Total patient visits and expected RA patients at each collaborating center
Approximate number of Anticipated number of
Country
RA patients enrolled in
clinic visits per year
the study per year
Jordan
2620
350 (13%)
Kingdom of Saudi Arabia
1500
100 ( 7%)
Lebanon
3000
300 (10%)
Qatar
3500
130 (4%)
United Arab Emirates
1600
250 (15%)
We are committed to 1000 patients and 1000 controls in the first year
Data collection
( 10-15 minutes)
• Clinical Data
– Demographic and clinical data questionnaire
– Ancestry questionnaire
• All data entry will be electronic via survey
gizmo
1. Demographic Questionnaire
2. Ethnicity Questionnaire
Blood Collection
(30 Minutes)
• Blood collection kits pre-labeled shipped to
center according to a preset schedule
• 10 cc of blood to be collected in EDTA tubes
• Tube to be inverted 8-10 times
• Blood to be transferred to cryovials, precoded
• Blood to be frozen immediately at -80 C
1. Shipment form
2. Blood collection form
Shipment
• Samples will then be shipped, on dry-ice,
according to a preset timeline, to WCMC-Q for
DNA extraction
Shipment lecture
Documentation
• Linking patients to samples
• Anonymization
• Recruitment strategy
WORKFLOW
Patient refuses to sign ICF
Eligibility Criteria met
Patient d/c
Patient signs two original
informed consent forms
and entered into the study
Pre-prepared labels affixed
to consent forms
•Use paper format
•Lable with provided lables
•Send electronically within 24
hours
Questionnaires filled
electronically by PI or
representative
Technical
problems
Patient to blood drawing
station
10 cc of blood drawn and
stored according to SOP
Blood shipped to WCMC-Q
per preset schedule
Patient refuses
blood draw
Document in log
sheet
Log sheet sent
monthly basis
Study Timeline
Aims
3/12-9/12 9/12-3/13
3/13-9/13
9/13-3/14
Set-up
Recruitmen
t
Famliy/exo
me
Genotype
Analysis
1000 patients
Sequencing
1000 patients
Analysis
3/14-9/14
9/14-3/15
Questions
Genetics of Rheumatoid Arthritis
in some Arab States
Thurayya Arayssi M.D
Weill Cornell Medical College-Qatar
May 2012
What is Next?
September 1 2012
First Patient to be recruited
June-September 2012
• Testing the system
– Use suvery Gizmo to trouble shoot
– Assess your site for readiness
• Identify responsibilities ( Responsibility log to be sent)
• -80 Freezer, racks, back up electricity and space
availability
• Pipettes
• Shipping agency and the requirements for shipping
blood outside the country
• Recruitment plan
June-September 2012
June and July
• SOP will be sent to each Center to standardize
and facilitate the work
• Weekly Skype calls in June to trouble shoot
according to a preset schedule
• Monthly Skype calls thereafter or as requested
by each center
June-September 2012
August
• Mid August– Reminder documents, webcasts sent to centers
– Skype calls for each Center to finalize needs prior
to “first patient in”
Communication Plans
• Monthly Skype Calls
• Quarterly Newsletter
• Update on recruitment
• Trouble shooting
• Emerging issues to all centers
• Meeting preset at the ACR and ? EULAR
Future Plans
• Searching for families with multiple members
with RA
• Metabolomics
THANK YOU