Laboratory for human genetics

Team: Lejla Kapur-Pojskić, PhD, Head of Laboratory; Naida Lojo-Kadrić, MSc; Jasmin Ramić; Ksenija Radić.

In order to establish a medical diagnosis, successful therapy and pharmacological treatment of many diseases it is often necessary to do genotyping of the disease. Contemporary methods of molecular genetics are also used worldwide in early evaluation of the effect of treatment or transplantations and they can even enable prediction of response to specific therapy.

Laboratory for Human Genetics has the capacity for characterization and genotyping of a wide range of inherited diseases. This includes the application of standard methods based on PCR. In families with high frequency of certain hereditary disorders it is possible to assess the risk for the future offspring or to do genotyping when there is medical indication.

Laboratory performs verified analytical methods such as:

- prenatal DNA characterization
- early determination of fetal sex and fatherhood
- PCR genotyping of HLA system
- detection of markers for different subtypes of acute myeloid (AML), acute lymphoblastic (ALL) and chronic myeloid leukemia (CML)
- neuromuscular dystrophies (Duchenne and Becker type)
-genotyping of the entire spectrum of inherited disorders through cooperation with laboratories specialized for the detection of rare hereditary disorders or diseases (www.genetests.org)

Laboratory for Human Genetics (LHG) was officially established by Internal act of the Board of the Institute for Genetic Engineering and Biotechnology in year 2004. Along with complementary laboratories of the Institute, LHG perfoms research and expert activities in the field of molecular-genetic characterization of human material.

In recent years, LHG  has successfully carried out 18 larger or smaller scientific projects related to the genetic characterization of human biological samples on the molecular-genetic level. It is particularly important to emphasize the results achieved in the field of strenghtening human resources and capacity of the laboratory through continuous education and activity supported by several funds for scientific research (Ministry of Education, Science, Culture and Sport of Sarajevo Canton and Federal Ministry of Education and Science, Federation of Bosnia and Herzegovina). One doctoral, three master and two diploma thesis within one international and three interinstitutional research scientific projects are being realized at the moment.

One of the strategic directions in the development of the Laboratory for Human Genetics is to strenghten the capacity for medical diagnosis. In that sense, we have established direct contact with the Clinical Centre of University in Sarajevo and Ministry of Health of Sarajevo Canton with the aim to clearly define the needs for medical diagnostics in Sarajevo Canton and Federation of Bosnia and Herzegovina.

A successful step forward in expanding the capacity of scientific research and cooperation has been made through the establishment of direct cooperation with the Institute of Oncology, Psychiatric Clinic and Hematological Clinic of University in Sarajevo. This has made the basis for the development and implementation of more complex scientific and research activities in the field of human genetics through a multidisciplinary approach and international participation. 

In the following period, projects of LHG will be focused on the further construction of new infrastructures and capacities, which will represent a basis for engagement in international networking and competitive collaborative research projects at a high research level.

 

Scientific activities

Overall scientific activities of the Laboratory during the past period can be classified into several areas:

Molecular-genetic characterization of hereditary mental disorders

Implementers: INGEB LHG, Psychiatric Clinic of KCUS

Mental disorders with proven hereditary etiology (bipolar disorder – BP1, schizophrenia, some anxiety disorders and disorders of neurodegenerative types etc.) represent complex genetic traits controlled by a huge number of genes which are also in complex interaction with environmental factors.

In the past decade a large number of gene loci which are associated with the aforementioned diseases has been identified as functional or positional candidates. On the global scale, identification of genes for bipolar disorder and schizophrenia is performed in terms of confirming the existing candidate genes as well as the locating new genes that are potentially associated with these disorders in larger samples of patients around the world.

Application of association analysis, i.e. linkage with the identified allelic variants of the applied markers with the occurrence of schizophrenia (χ2 or Fisher Exact test) showed statistically significant correlation at the level of P<0.05 for four observed markers (D5S818, 420M9-1395, D12S105 and RS5778313). 

Fisher Exact Test on the same level of reliability, along with the presence of the correction mechanism for a smaller statistical sample indicates a positive association of marker rs5778313 with schizophrenia. At the given level of statistical reliability higher than 99 % (P<0.01) for both tests, the positive association was found for marker D12S105 which is in the domain of DAAO gene located on chromosome 12. At the genotype level, positive association was confirmed for markers D12S105 and rs5778313; using the Fisher Exact correction factor it was confirmed only for the marker linked to the locus of DAAO with P<0.05.

Post-traumatic stress disorder (PTSD) in BiH represents a unique medical phenomenon in its etiology (trauma character). Current researches in the genetics of PTSD which have been performed in the LHG are aimed at the evaluation of association of genes which code dopaminergic factors (dopamine transporter 1 and dopamine receptor D2) with the appearance and character of PTSD in a sample of more than 200 examinees.

Bank of BiH population with more than 300 individual samples, i.e. 113 family triads has been established through cooperation of LHG with the Psychiatric Clinic of KCUS and german academic network for research of bipolar disorder. Distribution analysis of allelic and genotypic frequencies and their association with BP1 or particular symptoms of BP1 is planned in the further period.

 

Genotypic specificity of most common types of cancer in BiH in correlation with their phenotypic aspects

Implementers: LHG, Institut for Oncology of KCUS, Institut for Pathology of KCUS

Cancers are disorders with complex etiology with significant influence of genetic factors as the main trigger for processes of formation and development of tumor tissue. Within the project activities in this field it is noteworthy to set aside the results achieved so far, which are aimed at a comprehensive genetic characterization of established tumor bank and further development of biomedical research in BiH, as a basis for improvement of existing (advanced) medical and oncological practice in our country:

• Establishing a research base for the genetic characterization of the most common cancers in BiH (Phase I)
• Correlation of toxicity and levels of enzyme dihydropyrimidine dehydrogenase (DPD) and its genotype during the therapy with 5-fluorouracil
• Comparative analysis of difference between peripheral blood and bone marrow in diagnosis of BCR-ABL translocation using PCR method
• Coexpression analysis of proapoptotic factors (CDKN, CDK4, TP53 and BRCA1) in samples of tumor tissue
• Genotypic and phenotypic correlation of BRCA1 gene in a group of patients with breast and ovarian cancer.

 

Educational activities in the field of human molecular genetics and interdisciplinary biomedical researches

Implementers: LHG, University in Sarajevo

Since its constitution by the end of the academic year 2007/2008, LHG has participated in the practical and theoretical realization of many graduate and undergraduate works, as well as the diploma thesis, master thesis and doctoral dissertations.

Development of expertise in the field of medical genetics

Implementers: LHG, Hematological Clinic of KCUS

Within its capacities, from the beginning of 2004, LHG has been providing genetic testing of the presence of fusion gene BCR/ABL as a marker of chronic myeloid leukemia based on DNA analysis. The method that is used in the aforementioned analysis is based on reverse transcription of total mRNA isolated from the sample of bone marrow and peripheral blood of patient according to the method Chomczynski et Sacchi (1987) and target amplification using primers which encompass the fragment of the fusion gene. An internal protocol for monitoring minimal residual disease (MRD) based on Real-Time PCR has also been developed in the LHG. The next phase will include its validation on the tissue of patients.

Contact: lhg@ingeb.ba