# Impact of RNA splicing factor mutations on circular RNA biogenesis in leukemia

> **NIH NIH F31** · BECKMAN RESEARCH INSTITUTE/CITY OF HOPE · 2022 · $30,842

## Abstract

PROJECT SUMMARY/ABSTRACT
Approximately 10% of new cancer diagnoses in the United States are hematological malignancies that include
a spectrum of blood cancers and related disorders. In chronic lymphocytic leukemia (CLL) and myeloid
neoplasms, somatic hotspot mutations frequently occur in five RNA splicing factors (SFs): SF3B1, SRSF2,
U2AF1, ZRSR2, and genes encoding the U1 snRNA. These mutations drive aberrant splicing of mRNAs to
promote leukemogenesis. Recently, a novel class of RNAs called circular RNA (circRNA) was found to be
aberrantly expressed in many types of liquid tumors. Unlike mRNAs that form through normal splicing to produce
linear RNAs, circRNAs are produced through backsplicing that results in RNA circularization. Because of
advances to detection and annotation methods, circRNAs are now known to possess functional and clinical
significances, suggesting a novel role in cancer biology. However, to date, their precise role in hematological
malignancies, namely leukemia, remains undefined. The long-term goal is to investigate the functions and
therapeutic potentials of circRNAs. Moreover, the role of SF mutations (SF3B1, SRSF2, U2AF1, ZRSR2, and
genes encoding the U1 snRNA) in the aberrant expression of circRNAs remains unknown. Thus, the overall
objective is to link SF mutations to aberrant circRNA expressions. To this end, I hypothesize that mutations in
SF3B1 and other SFs such as SRSF2, U2AF1, ZRSR2, and genes encoding the U1 snRNA upregulate circRNA
abundance to promote leukemogenesis. The rationale for this research is that linking SF mutations to aberrant
expressions of circRNAs would define a novel regulatory axis, unlocking new therapeutic opportunities for
treating leukemia. Preliminary data from CLL patient B cells and cell lines showed that mutant SF3B1 promoted
aberrant expression of circRNAs in biological important molecular pathways such as protein transport and cell
cycle regulation. To identify circRNAs critical for cell survival, I have validated the emergent CRISPR CasRX
technology as a tool for screening circRNAs. I have also prototyped an experimental workflow for validating the
functions of these circRNAs. Additionally, to further determine the role of SF mutations on circRNA biogenesis,
cell lines with somatic mutations for SF3B1, SRSF2, U2AF1, ZRSR2, and genes encoding the U1 snRNA have
been established. Finally, I have engineered cell-based and minigene-based reporter systems to investigate the
mechanisms of backsplicing. Using this collection of tools, I propose the following aims: to identify and validate
the functional impact of SF mutation-associated circRNA in leukemia (Aim 1) and to determine the
mechanism of backsplicing (Aim 2). I expect the findings from this proposal will define a novel regulatory axis
linking SF mutations and circRNAs aberrant expressions to leukemogenesis.

## Key facts

- **NIH application ID:** 10464894
- **Project number:** 5F31CA261110-02
- **Recipient organization:** BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
- **Principal Investigator:** MIKE FERNANDEZ
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $30,842
- **Award type:** 5
- **Project period:** 2021-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10464894

## Citation

> US National Institutes of Health, RePORTER application 10464894, Impact of RNA splicing factor mutations on circular RNA biogenesis in leukemia (5F31CA261110-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10464894. Licensed CC0.

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