Genetics of Rheumatoid Arthritis in some Arab States

Thurayya Arayssi M.DWeill 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.

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.

Common Alleles Associated with RA Risk

T-Cell Activation NF Kappa B pathwayHLA DRB1:involved in MHC molecule–based antigen presentation and responsible for self-peptide selection and T-cell repertoire

REL:Proto-oncogene member of the NF-κB family that regulates leukocyte activation and survival

PTPN22:Lymphocyte-specific nonreceptor tyrosine phosphatase involved in regulation of activation threshold of lymphocyte

TNFAIP3:Signaling protein and negative regulator of TNF-α–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 pathwaysCD40:Costimulatory molecule that enhances interactions between T and B cells and increases auto- antibody production

BLK:B-lymphoid tyrosine kinase involved in B-cell receptor signaling and B-cell development

CTLA4:Costimulation suppressor that regulates interactions between T cells and antigen-presenting cells

CCL21

IL2RA:High-affinity receptor for interleukin-2 on lymphocyte subsets

FCGR2A

IL2:Cytokine that regulates activation of T cells, particularly regulatory T cells

PADI4:Enzyme that converts arginine to citrulline, creating autoantigens in rheumatoid arthritis

IL-21:Cytokine that regulates differentiation of T cells, particularly Th17, and activation of B cells

PRDM1

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

TNFRSF14

NEJM, 365;23nejm.orgdecember 8, 2011

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

ControlsInclusion 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 Country Approximate number of

clinic visits per yearAnticipated number of RA patients enrolled in 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 Questionnaire2. 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 form2. 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

Eligibility Criteria met

Patient signs two original informed consent forms

and entered into the study

Pre-prepared labels affixed to consent forms

Questionnaires filled electronically by PI or

representative

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 to sign ICF Patient d/c

Technical problems

•Use paper format•Lable with provided lables•Send electronically within 24 hours

Patient refuses blood draw

Document in log sheet

Log sheet sent monthly basis

WORKFLOW

Study TimelineAims 3/12-9/12 9/12-3/13 3/13-9/13 9/13-3/14 3/14-9/14 9/14-3/15

Set-upRecruitment

Famliy/exomeGenotype

Analysis

1000 patients 1000 patients

Sequencing Analysis

Questions

Genetics of Rheumatoid Arthritis in some Arab States

Thurayya Arayssi M.DWeill 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 2012June 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 2012August

• 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