Title: Clinical manifestation of hemophilia A in the absence of mutations in the F8 gene that encodes FVIII: role of microRNAs

Abstract

BACKGROUND Hemophilia A (HA) is associated with mutations in the F8 gene that expresses factor VIII (FVIII). Unexpectedly, HA also manifests in a small subset of individuals with no mutations (exonic or intronic) in their F8 gene. MicroRNAs (miRNAs) cause translational interference, affecting protein quality and stoichiometry. Here, by analyzing miRNAs of two patients from this subset, we evaluated miRNA‐based FVIII suppression as a testable hypothesis to explain FVIII deficiency in patients with HA with no F8 gene mutations. STUDY DESIGN AND METHODS To test the hypothesis, miRNA sequencing from two patients with mild and moderate HA with no mutations in their F8 gene, followed by experimental verification, was used to identify a group of upregulated miRNAs in patients with HA compared to normal controls; with binding sites in the 3′ untranslated region (UTR) of F8 messenger RNA (mRNA), a prerequisite for miRNA‐based gene regulation. From this pool, miR‐374b‐5p and miR‐30c‐5p, known to be expressed in human liver, where FVIII is expressed, were subjected to extensive characterization. RESULTS In two cell lines that constitutively express FVIII, we demonstrated that overexpression of miR‐374b or miR‐30c decreased FVIII expression, while an miR‐30c inhibitor partially restored FVIII expression. CONCLUSION These data support amore » role for microRNAs in fine‐tuning F8 gene regulation. Based on our findings, our current model suggests that in HA cases where the F8 gene is normal and is predicted to express normal levels of FVIII, F8 mRNA 3′ UTR targeting miRNAs may be responsible for a FVIII‐deficiency phenotype clinically manifesting as HA.« less

Authors:

Jankowska, Katarzyna I. ^[1]; McGill, Joseph ^[2]; Pezeshkpoor, Behnaz ^[3]; Oldenburg, Johannes ^[3]; Atreya, Chintamani D.  ^[1];

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Sauna, Zuben E. ^[2]

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+ Show Author Affiliations

1. OBRR/DBCD/LCH US Food and Drug Administration Silver Spring Maryland
2. OTAT/DPPT/HB in the Center for Biologics Evaluation and Research US Food and Drug Administration Silver Spring Maryland
3. Institute of Experimental Hematology and Transfusion Medicine University Clinic Bonn Bonn Germany, Center for Rare Diseases Bonn (ZSEB) University Clinic Bonn Bonn Germany

Publication Date:

Fri Nov 29 00:00:00 EST 2019

Sponsoring Org.:

USDOE

OSTI Identifier:

1576660

Resource Type:

Publisher's Accepted Manuscript

Journal Name:

Transfusion (Philadelphia)

Additional Journal Information:

Journal Name: Transfusion (Philadelphia) Journal Volume: 60 Journal Issue: 2; Journal ID: ISSN 0041-1132

Publisher:

Wiley-Blackwell

Country of Publication:

United States

Language:

English