Further Evidence That MicroRNAs Can Play a Role in Hemophilia A Disease Manifestation: F8 Gene Downregulation by miR-19b-3p and miR-186-5p

Abstract

Hemophilia A (HA) is a F8 gene mutational disorder resulting in deficiency or dysfunctional FVIII protein. However, surprisingly, in few cases, HA is manifested even without mutations in F8 . To understand this anomaly, we recently sequenced microRNAs (miRNAs) of two patients with mild and moderate HA with no F8 gene mutations and selected two highly expressing miRNAs, miR-374b-5p and miR-30c-5p, from the pool to explain the FVIII deficiency that could be mediated by miRNA-based F8 /FVIII suppression. In this report, an established orthogonal in vivo RNA-affinity purification approach was utilized to directly identify a group of F8 -interacting miRNAs and we tested them for F8 /FVIII suppression. From this pool, two miRNAs, miR-19b-3p and miR-186-5p, were found to be upregulated in a severe HA patient with a mutation in the F8 coding sequence and two HA patients without mutations in the F8 coding sequence were selected to demonstrate their role in F8 gene expression regulation in mammalian cells. Overall, these results provide further evidence for the hypothesis that by targeting the 3′UTR of F8 , miRNAs can modulate FVIII protein levels. This mechanism could either be the primary cause of HA in patients who lack F8 mutations or control the severity of the disease in patients with F8 mutations.

RNA sequencing suggests that non‐coding RNAs play a role in the development of acquired haemophilia

“…Thus, the interaction between non‐coding RNAs and F8 gene might represent an important path in understanding how patients without F8 mutations are lacking FVIII in blood and develop HA. It was shown that miR‐374b‐5p and miR‐30c‐5b target F8 gene and impair FVIII, indicating that classical HA could be evaluate not only by the mutational status 5,21 …”

Effects of Shear Stress on Production of FVIII and vWF in a Cell-Based Therapeutic for Hemophilia A

“…A recent report of HA in human patients who lack F8 mutations, showed that overexpression of miR-30c decreased FVIII expression, while a miR-30c inhibitor partially restored FVIII expression in two cell lines that constitutively express FVIII ( Jankowska et al, 2020a ). These findings led the authors to hypothesize that expression of miRNAs targeting the 3’-UTR of FVIII mRNA can modulate FVIII levels and may be responsible for a FVIII-deficiency phenotype that clinically manifests as HA ( Jankowska et al, 2020b ). Because KLF2 overexpression has been shown to induce upregulation of the miR-30 family members miR 30b and 30c ( Doebele et al, 2018 ), we next examined the levels of miR30c in PLC-mcoET3 in static vs. flow conditions and demonstrated that exposure to flow led to a nearly threefold induction of miR-30c in PLC-mcoET3, providing a mechanistic explanation for the differential response of the endogenous/native FVIII and the mcoET3 transgene to conditions of flow.…”

Dysregulated miRNAs network in the critical COVID-19: An important clue for uncontrolled immunothrombosis/thromboinflammation

“…by targeting the factor VIII (FVIII), and miR-18a, miR-29a/b/c, miR-211, miR-193-3p, miR-186, etc. by targeting the fibrinogen (Fg) regulate the common coagulation pathway [202] , [203] , [204] , [205] ( Fig. 2 b-B1-3, 2b-D).…”