LIFE SCIENCE RESEARCH SEMINARS
Web: http://www.um.edu.mt/events/scisem/ Email: scisem@um.edu.mt
Abstract form
Title:
Models to Understand the Developmental Control of Foetal
Haemoglobin
Presenter:
Laura Grech
Contact address:
Tel:
79959025
Fax:
Email: lauragrech@yahoo.com
Presentation date:
7 th April 2014
Abstract
In this study we used three different available models to study the developmental control and regulation of HbF. These models included Haemoglobin F Malta I heterozygote (cords n=200; adults n=20), HPFH-KLF1 haploinsufficient family members (n=5) and Cell Models (primary human erythroid progenitor cells) (n=15). PCR genotyping for XMN1, BCL11A and HBS1L-MYB and
DNA sequencing for (AT)xTy and KLF1 was carried out on all samples. Erythroid progenitor cells were cultured from 15 donors (5 healthy adults, 5 thalassaemic homozygotes and 5 KLF1 haploinsufficient family members). Proliferating and differentiating cells were cultured under the effect of Thalidomide, 5’-Aza-2’-deoxycytidine and Hydroxyurea. The cell pellets extracted from these cultures on different culturing days were used for globin chain analysis and for RNA expression of globin and other trans -acting genes. When real-time qPCR was performed, an increa se in gamma globin and downregu l ation of KLF1 in the HEP s treated with 1μM thalidomide was noted in one thalassaemic homozygote patient carrying the IVSI-6C/COD39T mutation. An increase in gamma globin and decrease in KLF1 was also noted in three out of four t halassaemic patients. With hydro xyurea 10μM and 100μM the least responsive thalass a emic patient was the one carrying the long isoform of (AT)
X
T
Y
that is the (AT)
9
T
5
. All the other thalassaemic patients carried the short isoform of the (AT)
X
T
Y
that is the (AT)
7
T
7
. It is known that disruptive ATxTy (long form) give rise to an increase in β-globin bec ause BP1 is inhibited from binding. The data of all three models will be presented and discussed regarding to their connection on HbF regulation and control.