Department of Hematogenetics
Projects
Completed Projects
1. Molecular Genetics of major Hemoglobinopathies in India
Funding Agency: Indo – French Centre for promotion of Advanced Research, New Delhi
Duration : 3 years (1992-1995)
Outcome: This collaborative programme with Institute of Pathologic Moleculaire, Paris enabled us to establish several molecular techniques at the Institute. The molecular genetics of sickle cell disease in a tribal population from Gujarat and a non tribal population from Maharashtra was studied by undertaking haplotype analysis, a genotying by Southern blotting and the Xmn1 polymorphism analysis. The Denaturing Gradient Gel Electrophoresis technique was established and used to localize b-thalassemia mutations and identify them by DNA sequencing.
2. Thalassemia in the Indian sub-continent – Prevention and Clinical Management
Funding Agency : INSERM, Paris
Duration: 2 years (1995-1997)
Outcome: Covalent Reverse Dot Blot Hybridization was established for screening for 6 Indian b-thalassemia mutations along with Hb S and Hb E. b-thalassemia mutations, a genotype and the Xmn1 polymorphism were studied in patients with b-thalassemia major and intermedia to evaluate their effect of an the clinical phenotype of the disease.
3. R&D Cum Genetic Clinic for Molecular Diagnosis of inherited Hematological Disorders
Funding Agency : Dept. of Biotechnology, New Delhi
Duration: 2 years (1997-1999)
Outcome: A diagnostic kit was prepared for molecular characterization of 6 common Indian b-thalassemia mutations along with Hb S and E and patented with patent office, Govt. of India. Using this along with other techniques like ARMS, DGGE and DNA sequencing, b-thalassemia mutations were characterized and first trimester prenatal diagnosis done for couples at risk of thalassemia and sickle cell disease.
4. Jai Vigyan S & T Mission project on Community Control of Thalassemia: Awareness, Screening, Genetic Counselling and Prevention
Funding Agency: ICMR
Duration: 5 years (2000-2005)
Outcome: In this multicentric project co-ordinated by us included six centers in different states of India ( Punjab, Asam, W.Bengal, Karnataka, Gujarat and Maharashtra) where awareness on the thalassemias was generated among college students and women attending antenatal clinics. 60,000 individuals from these two groups were screened using both single approaches like one tube osmotic fragility as well as by suing RBC indices and HPLC analysis to evaluate the usefulness of different techniques and to determine the prevalence of hemoglobinopathies in different regions. Genetic counseling was given and couples at risk sent to our Institute for prenatal diagnosis whenever possible.
5. Intervention Programme for Nutritional Anemia and hemoglobinopathies among some primitive tribal populations of India
Funding Agency: ICMR
Duration: 6 years (1999-2005)
Outcome: This was a multicentric project co-ordinated by us and undertaken in 4 states (Tamil nadu, Maharashtra, Gujarat and Orissa) for clinical evaluation and screening for nutritional anemia, sickle cell disease and other hemoglobinopathies as well as G6PD deficient among primitive tribal populations in these regions. The prevalence of nutritional anemia and sickle cell trait and disease was established and intervention given to individuals with iron deficiency and sickle cell disease and these cases were followed up to evaluate the effectiveness of the intervention programme.
6. Community Control Programme on hemoglobinopathies
Funding Agency: WHO-ICMR
Duration: 2 years (2002-2004)
Outcome: This was a pilot study co-ordinated by us in 3 states Chattisgarh, Madhya Pradesh and Bihar where training was given to Medical Officers and Laboratory Technicians from these states at a workshop conducted at Raipur for identification and management of hemoglobinopathies. Preliminary screening for hemoglobinopathies was then done at Raipus, Jabalpur and Patna with confirmation of the results at our Institute to know the frequencies of hemoglobinopathies in these states.
7. Incidence and Molecular Characterization of G6PD deficiency in North East India
Funding Agency: ICMR
Duration: 3 years (2003-2006)
Outcome: This was a collaborative project with RMRC, Dibrugarh. Screening for G6PD deficiency was done in the North Eastern region mainly in Assam and Mizoram. The G6PD deficient samples were sent to our Institute for characterization of the mutations.
8. Training Centre for Molecular Genetics, Cytogenetics and Cell biology - Application in Hematological disorders
Funding Agency: Dept. of Biotechnology, New Delhi
Duration: 2 years (2005-2007)
Outcome: Hands on training in molecular diagnosis of thalassemias and other hemoglobinopathies was given in our department while training in molecular diagnosis of hemophilia and molecular cytogenetic analysis was given in the Department of Hemostasis and Cytogenetics respectively. Both medical and non-medical scientists and technologists from different parts of the country participated in the 3 week training programme. In the second year individual scientists/technologists came for a longer period (1 month to 3 months) for specialization in their area of interest.
9. Glucose-6-Phosphate dehydrogenase mutations and haplotypes in various ethnic groups of India .
Funding agency : ICMR
Duration: 2004 – 2008
Outcome: A total of seven mutations in exon 3,4,6,7 and 9 of the G6PD gene were identified which include a novel and two very rare and unreported variants from India .
This study also indicates the genetic heterogeneity of G6PD mutations as well as the segregation of these mutations among the tribal groups of India .
G6PD Orissa (131 C→ G) and G6PD Namoru (208 T→C) mutations are presumed to be older than the others since it is mainly found in the tribal populations which are considered to be the original inhabitants of India .
10. Molecular pathology of some hematological disorders
Funding Agency: ICMR
Duration: 3 years (2006-2009)
Outcome: There were 4 projects undertaken by us under this programme.
a. Non-invasive prenatal diagnosis of hemoglobinopathies from maternal blood
Fetal NRBCs were isolated from maternal blood by enrichment followed by flow sorting using different monoclonal antibodies and the diagnosis was done using a nested PCR approach. The success rate for an accurate diagnosis was 85-90%.
Real time PCR based approaches were developed for fetal diagnosis using cell free fetal DNA from maternal plasma.
b. The effect of hydroxyurea on the fetal hemoglobin response and clinical response in sickle cell disease and the b-thalassemia
Hydroxyurea was started in pediatric and adult patients with sickle cell disease and sickle thalassemia having severe clinical manifestations and initially high Hb F levels. Genetic analysis included haplotyping, a genotyping and Xmn1 polymorphism analysis. The patients were followed up haemtologically and clinically for 2 years and these findings were correlated with the genetic parameters in the responders and non-responders. Similar studies were undertaken in thalassemia intermedia, thalassemia major and Hb E thalassemia patients
c. Non deletional a thalassemia in Indians
Individuals suspected to have a thalassemia based on hematological analysis and Hb electrophoresis/HPLC analysis were first screened for a gene deletions using multiplex PCR. When these were absent CSGE analysis followed by DNA sequencing was done to look for non - deletional a thalassemia or other a chain variants. Five unusual a chain varints were identified.
d. Gene expression studies in the thalassemia
Real Time PCR was used to look at the a, b, g, & dmRNA expression in individuals with different globin gene mutations. The meon b/a synthetic ratio was much lower in b-thalassemia major patients as against thalassemia intermedia patients. Quantification of d globin mRNA levels in b thalassemia trait suggested that the increase in Hb A 2 levels was due to an increase in the transcriptional activity of d globin gene.
Ongoing Projects
1. Community Control of Thalassemia – Establishment of molecular characterization of hemoglobinopathies and prenatal diagnosis of thalassemia and sickle cell disease at regional centers
Funding Agency: ICMR
Duration: 3 years (2008-2011)
Expected Outcome: This is the second phase of the earlier multicentric Jai Vigyan Project where awareness on thalassemias and other hemoglobinopathies was generated and diagnostic centers established in different regions. In this project, 5 of the earlier centers have been selected in Gujarat, Maharashtra, Karntaka, W.Bengal and Punjab where we will establish molecular analysis and prenatal diagnosis.
2. Molecular Characterization of hyperbilirubinemia among the neonates in India
Funding Agency: Dept. of Biotechnology, New Delhi
Duration: 3 years (2009-2012)
Expected Outcome: Molecular characterization of different genes (G6PD, UGT!A!, OATP2) associated with hyperbilirubinemia may help us to understand the genetic basis of hyperbilirubinemia in neonates.
3. Indo-US project - Pilot study on newborn screening for hemoglobinopathies in South Gujarat, India
Funding Agency: ICMR & National Institutes of Health, USA
Duration : 2008 – 2010
Expected Outcome : This project will help to evaluate the feasibility of undertaking newborn screening for sickle cell disease in a tribal area in South Gujarat and to know the true clinical severity of the disease in these tribal groups by raising a cohort and following the cases regularly to offer early comprehensive care. This will also serve as a model for extending newborn screening to other tribal areas in the country.
4. Assessment of neonatal screening approaches for sickle cell disease and the effect of early intervention in management of the disease
Funding agency: Dept. of Biotechnology, New Delhi
Duration: 2009- 2012
Expected Outcome: This collaborative project with Govt.Medical College, Nagpur will help to assess the most feasible and cost effective approach for newborn screening for sickle cell disorders using a hospital based targeted screening approach where babies of sickle heterozygous mothers are screened in mainly a non-tribal population. A cohort of 100 sickle homozygous babies will be registered at the sickle cell clinic and follow up regularly with timely intervention and management of complications. The effect of genetic modifiers on the time of presentation and severity of the disease will also be assessed.
5. Workshops on Diagnosis and Molecular Characterization of Hemoglobinopathies
Duration - 2010-2012
Expected Outcome: This Translational Research activity aims at conducting wet workshops for medical colleges in different regions of the country to familiarize them with the molecular techniques used for characterization of b-thalassemias and other hemoglobinopathies.
6. Molecular Characterization of Hyperbilirubinemia among the neonates in India
Funding Agency: Dept. of Biotechnology, New Delhi
Duration: 3 years (2009-2011)
Expected Outcome:
A total of seven SNPs in the promoter and coding region of the UGTIAI gene were studied. Of these, 3 were non polymorphic in our population. Promoter polymorphisms were found to have a significant effect on bilirubin concentrations. Presence of mutant alleles either in homozygous or heterozygous condition had a significant effect on the development of neonatal hyperbilrubinemia.
7. Understanding the unexplained hyperbilirubinemia in hemoglobinopathies
Funding Agency: ICMR Extramural grant
Duration: 3 years (2010-2012)
Expected Outcome:
This study will help to determine the effect of polymorphic variants of different genes associated with hyperbilirubinemia in patients with hemoglobinopathies as well as normal control groups and subsequently to correlate the genotypic and phenotypic expression of these cases
8. Evaluation of Alpha Hemoglobin Stabilizing Protein (AHSP) as a genetic modifier and effect of linked and unlinked determinants on clinical manifestations of Hemoglobinopathies:
Funding agency : ICMR
Duration : 3 years (2009-2012)
Expected Outcome : This study will help us to find out the role of different primary and secondary genetic modifiers in clinical manifestations of the disease. It will also help us to know the role of AHSP as a genetic modifier of thalassemia. Knowing the role of different genetic markers in clinical manifestations will ultimately help us in giving genetic counseling and prenatal diagnosis to the couple at risk
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9. Genotyping of subjects with borderline haemoglobin A 2 levels and effect of linked and unlinked determinants on β thalassemia carrier screening.
Funding agency : DBT
Duration : 3years (2011-2014)
Expected Outcome : A simple cost effective approach for genotyping of subjects with borderline haemoglobin A 2 levels will be established and the effect of linked and unlinked determinants on β thalassaemia carrier screening will also be studied in detail. The benefits of identifying silent gene b thalassemia individuals will help in reducing the birth of homozygous children. Prenatal diagnosis offered to couples at-risk will reduce the health burden of β thalassemia in the country.
10. Modulation of the Clinical Phenotypes of Sickle Cell Disease by Genetic Polymorphisms in Modifier Genes.
Funding agency : ICMR
Duration : 3 years (2011-2014)
Expected Outcome : This study will help us to know the multigenic contribution towards clinical variability in Indian patients with sickle cell disease.It will also help us to determine the effect of different linked and unlinked genetic modifiers on specific sub-phenotypes in these patients.Knowing the role of different genetic markers in clinical manifestations of the disease will ultimately help us in better management of the patients .
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