Small ncRNA expression-profiling of blood from hemophilia a patients identifies mir-1246 as a potential regulator of Factor 8 gene

Tewarit Sarachana, Neetu Dahiya, Vijaya L. Simhadri, Gouri Shankar Pandey, Surbhi Saini, Christine Guelcher, Michael F. Guerrera, Chava Kimchi-Sarfaty, Zuben E. Sauna, Chintamani D. Atreya

Research output: Contribution to journalArticle

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Abstract

Hemophilia A (HA) is a bleeding disorder caused by deficiency of functional plasma clotting factor VIII (FVIII). Genetic mutations in the gene encoding FVIII (F8) have been extensively studied. Over a thousand different mutations have been reported in the F8 gene. These span a diverse range of mutation types, namely, missense, splice-site, deletions of single and multiple exons, inversions, etc. There is nonetheless evidence that other molecular mechanisms, in addition to mutations in the gene encoding the FVIII protein, may be involved in the pathobiology of HA. In this study, global small ncRNA expression profiling analysis of whole blood from HA patients, and controls, was performed using high-throughput ncRNA microarrays. Patients were further sub-divided into those that developed neutralizing- anti-FVIII antibodies (inhibitors) and those that did not. Selected differentially expressed ncRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. We identified several ncRNAs, and among them hsa-miR- 1246 was significantly up-regulated in HA patients. In addition, miR-1246 showed a six-fold higher expression in HA patients without inhibitors. We have identified an miR-1246 target site in the noncoding region of F8 mRNA and were able to confirm the suppressory role of hsa-miR-1246 on F8 expression in a stable lymphoblastoid cell line expressing FVIII. These findings suggest several testable hypotheses vis-à-vis the role of nc-RNAs in the regulation of F8 expression. These hypotheses have not been exhaustively tested in this study as they require carefully curated clinical samples.

Original languageEnglish (US)
Article numbere0132433
JournalPloS one
Volume10
Issue number7
DOIs
StatePublished - Jul 15 2015

Fingerprint

hemophilia
factor VIII
Factor VIII
Hemophilia A
Blood
Genes
blood
mutation
Gene encoding
genes
Mutation
blood coagulation factors
Blood Coagulation Factors
Polymerase chain reaction
Missense Mutation
Transcription
Microarrays
neutralization
Reverse Transcription
exons

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Sarachana, T., Dahiya, N., Simhadri, V. L., Pandey, G. S., Saini, S., Guelcher, C., ... Atreya, C. D. (2015). Small ncRNA expression-profiling of blood from hemophilia a patients identifies mir-1246 as a potential regulator of Factor 8 gene. PloS one, 10(7), [e0132433]. https://doi.org/10.1371/journal.pone.0132433
Sarachana, Tewarit ; Dahiya, Neetu ; Simhadri, Vijaya L. ; Pandey, Gouri Shankar ; Saini, Surbhi ; Guelcher, Christine ; Guerrera, Michael F. ; Kimchi-Sarfaty, Chava ; Sauna, Zuben E. ; Atreya, Chintamani D. / Small ncRNA expression-profiling of blood from hemophilia a patients identifies mir-1246 as a potential regulator of Factor 8 gene. In: PloS one. 2015 ; Vol. 10, No. 7.
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abstract = "Hemophilia A (HA) is a bleeding disorder caused by deficiency of functional plasma clotting factor VIII (FVIII). Genetic mutations in the gene encoding FVIII (F8) have been extensively studied. Over a thousand different mutations have been reported in the F8 gene. These span a diverse range of mutation types, namely, missense, splice-site, deletions of single and multiple exons, inversions, etc. There is nonetheless evidence that other molecular mechanisms, in addition to mutations in the gene encoding the FVIII protein, may be involved in the pathobiology of HA. In this study, global small ncRNA expression profiling analysis of whole blood from HA patients, and controls, was performed using high-throughput ncRNA microarrays. Patients were further sub-divided into those that developed neutralizing- anti-FVIII antibodies (inhibitors) and those that did not. Selected differentially expressed ncRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. We identified several ncRNAs, and among them hsa-miR- 1246 was significantly up-regulated in HA patients. In addition, miR-1246 showed a six-fold higher expression in HA patients without inhibitors. We have identified an miR-1246 target site in the noncoding region of F8 mRNA and were able to confirm the suppressory role of hsa-miR-1246 on F8 expression in a stable lymphoblastoid cell line expressing FVIII. These findings suggest several testable hypotheses vis-{\`a}-vis the role of nc-RNAs in the regulation of F8 expression. These hypotheses have not been exhaustively tested in this study as they require carefully curated clinical samples.",
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Sarachana, T, Dahiya, N, Simhadri, VL, Pandey, GS, Saini, S, Guelcher, C, Guerrera, MF, Kimchi-Sarfaty, C, Sauna, ZE & Atreya, CD 2015, 'Small ncRNA expression-profiling of blood from hemophilia a patients identifies mir-1246 as a potential regulator of Factor 8 gene', PloS one, vol. 10, no. 7, e0132433. https://doi.org/10.1371/journal.pone.0132433

Small ncRNA expression-profiling of blood from hemophilia a patients identifies mir-1246 as a potential regulator of Factor 8 gene. / Sarachana, Tewarit; Dahiya, Neetu; Simhadri, Vijaya L.; Pandey, Gouri Shankar; Saini, Surbhi; Guelcher, Christine; Guerrera, Michael F.; Kimchi-Sarfaty, Chava; Sauna, Zuben E.; Atreya, Chintamani D.

In: PloS one, Vol. 10, No. 7, e0132433, 15.07.2015.

Research output: Contribution to journalArticle

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T1 - Small ncRNA expression-profiling of blood from hemophilia a patients identifies mir-1246 as a potential regulator of Factor 8 gene

AU - Sarachana, Tewarit

AU - Dahiya, Neetu

AU - Simhadri, Vijaya L.

AU - Pandey, Gouri Shankar

AU - Saini, Surbhi

AU - Guelcher, Christine

AU - Guerrera, Michael F.

AU - Kimchi-Sarfaty, Chava

AU - Sauna, Zuben E.

AU - Atreya, Chintamani D.

PY - 2015/7/15

Y1 - 2015/7/15

N2 - Hemophilia A (HA) is a bleeding disorder caused by deficiency of functional plasma clotting factor VIII (FVIII). Genetic mutations in the gene encoding FVIII (F8) have been extensively studied. Over a thousand different mutations have been reported in the F8 gene. These span a diverse range of mutation types, namely, missense, splice-site, deletions of single and multiple exons, inversions, etc. There is nonetheless evidence that other molecular mechanisms, in addition to mutations in the gene encoding the FVIII protein, may be involved in the pathobiology of HA. In this study, global small ncRNA expression profiling analysis of whole blood from HA patients, and controls, was performed using high-throughput ncRNA microarrays. Patients were further sub-divided into those that developed neutralizing- anti-FVIII antibodies (inhibitors) and those that did not. Selected differentially expressed ncRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. We identified several ncRNAs, and among them hsa-miR- 1246 was significantly up-regulated in HA patients. In addition, miR-1246 showed a six-fold higher expression in HA patients without inhibitors. We have identified an miR-1246 target site in the noncoding region of F8 mRNA and were able to confirm the suppressory role of hsa-miR-1246 on F8 expression in a stable lymphoblastoid cell line expressing FVIII. These findings suggest several testable hypotheses vis-à-vis the role of nc-RNAs in the regulation of F8 expression. These hypotheses have not been exhaustively tested in this study as they require carefully curated clinical samples.

AB - Hemophilia A (HA) is a bleeding disorder caused by deficiency of functional plasma clotting factor VIII (FVIII). Genetic mutations in the gene encoding FVIII (F8) have been extensively studied. Over a thousand different mutations have been reported in the F8 gene. These span a diverse range of mutation types, namely, missense, splice-site, deletions of single and multiple exons, inversions, etc. There is nonetheless evidence that other molecular mechanisms, in addition to mutations in the gene encoding the FVIII protein, may be involved in the pathobiology of HA. In this study, global small ncRNA expression profiling analysis of whole blood from HA patients, and controls, was performed using high-throughput ncRNA microarrays. Patients were further sub-divided into those that developed neutralizing- anti-FVIII antibodies (inhibitors) and those that did not. Selected differentially expressed ncRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. We identified several ncRNAs, and among them hsa-miR- 1246 was significantly up-regulated in HA patients. In addition, miR-1246 showed a six-fold higher expression in HA patients without inhibitors. We have identified an miR-1246 target site in the noncoding region of F8 mRNA and were able to confirm the suppressory role of hsa-miR-1246 on F8 expression in a stable lymphoblastoid cell line expressing FVIII. These findings suggest several testable hypotheses vis-à-vis the role of nc-RNAs in the regulation of F8 expression. These hypotheses have not been exhaustively tested in this study as they require carefully curated clinical samples.

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