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THE HIF1A/2A MRNA INDEX HAS A SIMILAR TREND AS THE CHANGES OF EXPRESSION MRNA CALR AND MDR1 GENES IN WHOLE BLOOD SAMPLES OF PATIENTS WITH JAK2 V617F POSITIVE MPN
Author(s):
Aleksey Gorbenko
,
Aleksey Gorbenko
Affiliations:
Department of Health,Krasnoyarsk branch of the Federal State budgetary Institution «Hematology Research Center»,Krasnoyarsk,Russian Federation
Marina Stolyar
,
Marina Stolyar
Affiliations:
Department of Health,Krasnoyarsk branch of the Federal State budgetary Institution «Hematology Research Center»,Krasnoyarsk,Russian Federation;Siberian Federal University,Krasnoyarsk,Russian Federation
Mikhail Mikhalev
,
Mikhail Mikhalev
Affiliations:
Department of Health,Krasnoyarsk branch of the Federal State budgetary Institution «Hematology Research Center»,Krasnoyarsk,Russian Federation;Krasnoyarsk сity Clinical Hospital № 7,Krasnoyarsk,Russian Federation
Evgeniy Vasiliev
,
Evgeniy Vasiliev
Affiliations:
Department of Health,Krasnoyarsk branch of the Federal State budgetary Institution «Hematology Research Center»,Krasnoyarsk,Russian Federation;Krasnoyarsk regional hospital,Krasnoyarsk,Russian Federation
Vladimir Khorzhevskyi
,
Vladimir Khorzhevskyi
Affiliations:
Krasnoyarsk State Regional Bureau of Pathology, Krasnoyarsk,Russian Federation
Igor Olkhovskiy
Igor Olkhovskiy
Affiliations:
Department of Health,Krasnoyarsk branch of the Federal State budgetary Institution «Hematology Research Center»,Krasnoyarsk,Russian Federation;Krasnoyarsk Scientific Center SB RAS,Krasnoyarsk,Russian Federation
(Abstract release date: 05/18/17) EHA Library. Stolyar M. 05/18/17; 182742; PB2028
Marina Stolyar
Marina Stolyar
Contributions
PDF Abstract
Abstract

Abstract: PB2028

Type: Publication Only

Background

Various groups have reported that isoforms of hypoxia-inducible transcription factor 1a (HIF-1a) and 2a (HIF-2a) can regulate both overlapping and distinct target genes. HIF-1a and HIF-2a have been shown to play opposite roles in the regulation of macrophage function [Takeda N. e.a., 2010]. HIF-index incorporated as a strong prognostic biomarker of renal cell cancer [Szendrői A. e.a., 2016]. Only HIF1a was known as regulator expression of multidrug resistance gene (MDR1) and response to chemotherapy [Comerford K.M. e.a., 2002]. New data have shown exclusive role of HIF-2α in regulates the proliferation and glucose metabolism in erythroleukemia cells [Xu Q.Q. e.a., 2016]. No any information about relations between index of HIF and mRNA gene expression level MDR1 or CALR in patients with myeloproliferative neoplasms (MPN).

Aims
Investigate the mRNA expression levels of HIF-1a and HIF-2a, MDR1 and CALR genes in whole blood samples from patients with JAK2 V617F positive MPN.

Methods
Real-time PCR was performed to detect of HIF1a, HIF2a, MDR1 and CALR mRNA transcripts levels in white blood cells 14 healthy volunteers (median age 22 years, range 21-58 years, 57% males) and 11 (median age 44 years, range 20-77 years, 45% males) patients with JAK2 V617F-positive MPN, median of allelic burden is 36%, range 9-87%. Venous blood were collected in tube with RNAse inhibitor. Total RNA was isolated using “RIBO-zol-D” (Aplisens) and were transcribed using “Reverta-L” (Aplisens). PCR was optimized for the thermocycler CFX96 (Bio-Rad). The results were calculated utilizing the delta Ct method in the software package of “R”. The threshold cycles (Ct) genes and housekeeping genes (TBP, GUS, ABL) determined using Cy0 method. The results was normalization with this reference genes. Mann-Whitney U test was used to evaluate significant difference between the groups, the degree of correlation (r) was assessed using Spearman test.

Results

We observed a lower mRNA expression MDR1 and CALR in whole blood samples of patients with MPN compared with a group of healthy volunteers (fig. 1). The expression level of mRNA HIF2a not changed and for HIF1a it should be noted a tendency for statistical significance. It found no correlation between allelic burden and mRNA expression level. Index HIF 1a/2a more clearly showed a correlation with the fall of MDR1 and CALR mRNAs (r= -0.64 in control and r= -0.7 in MPN group, p<0.05). CALR gene unlike MDR1 gene is not known among the target HIF regulation, but their unidirectional change indicates the possible metabolic links.

Conclusion

We assume that the studied gene expression changes reflect the metabolic processes in the bone marrow progenitor cells. Probably JAK2 V617F mutation leads to more favorable microenvironment and reduced willingness to autophagy, causing the index shift HIF1a/2a. We found reduced of mRNA CALR expression in blood cells at MPN and this fact require further investigation.

Session topic: 15. Myeloproliferative neoplasms - Biology

Keyword(s): Myeloproliferative disorder, MDR1, Gene expression

Abstract: PB2028

Type: Publication Only

Background

Various groups have reported that isoforms of hypoxia-inducible transcription factor 1a (HIF-1a) and 2a (HIF-2a) can regulate both overlapping and distinct target genes. HIF-1a and HIF-2a have been shown to play opposite roles in the regulation of macrophage function [Takeda N. e.a., 2010]. HIF-index incorporated as a strong prognostic biomarker of renal cell cancer [Szendrői A. e.a., 2016]. Only HIF1a was known as regulator expression of multidrug resistance gene (MDR1) and response to chemotherapy [Comerford K.M. e.a., 2002]. New data have shown exclusive role of HIF-2α in regulates the proliferation and glucose metabolism in erythroleukemia cells [Xu Q.Q. e.a., 2016]. No any information about relations between index of HIF and mRNA gene expression level MDR1 or CALR in patients with myeloproliferative neoplasms (MPN).

Aims
Investigate the mRNA expression levels of HIF-1a and HIF-2a, MDR1 and CALR genes in whole blood samples from patients with JAK2 V617F positive MPN.

Methods
Real-time PCR was performed to detect of HIF1a, HIF2a, MDR1 and CALR mRNA transcripts levels in white blood cells 14 healthy volunteers (median age 22 years, range 21-58 years, 57% males) and 11 (median age 44 years, range 20-77 years, 45% males) patients with JAK2 V617F-positive MPN, median of allelic burden is 36%, range 9-87%. Venous blood were collected in tube with RNAse inhibitor. Total RNA was isolated using “RIBO-zol-D” (Aplisens) and were transcribed using “Reverta-L” (Aplisens). PCR was optimized for the thermocycler CFX96 (Bio-Rad). The results were calculated utilizing the delta Ct method in the software package of “R”. The threshold cycles (Ct) genes and housekeeping genes (TBP, GUS, ABL) determined using Cy0 method. The results was normalization with this reference genes. Mann-Whitney U test was used to evaluate significant difference between the groups, the degree of correlation (r) was assessed using Spearman test.

Results

We observed a lower mRNA expression MDR1 and CALR in whole blood samples of patients with MPN compared with a group of healthy volunteers (fig. 1). The expression level of mRNA HIF2a not changed and for HIF1a it should be noted a tendency for statistical significance. It found no correlation between allelic burden and mRNA expression level. Index HIF 1a/2a more clearly showed a correlation with the fall of MDR1 and CALR mRNAs (r= -0.64 in control and r= -0.7 in MPN group, p<0.05). CALR gene unlike MDR1 gene is not known among the target HIF regulation, but their unidirectional change indicates the possible metabolic links.

Conclusion

We assume that the studied gene expression changes reflect the metabolic processes in the bone marrow progenitor cells. Probably JAK2 V617F mutation leads to more favorable microenvironment and reduced willingness to autophagy, causing the index shift HIF1a/2a. We found reduced of mRNA CALR expression in blood cells at MPN and this fact require further investigation.

Session topic: 15. Myeloproliferative neoplasms - Biology

Keyword(s): Myeloproliferative disorder, MDR1, Gene expression

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