Decoding ALS: How Environmental Exposure Alters the RNA Pseudouridylation Landscape and RNA-protein Interactions

Environmental exposure can alter pseudouridine (Ψ) landscape in cellular RNA. Ψ is the most abundant modified nucleoside in nature and it regulates many RNA processes. Similar to N6-methyladenosine (m6A), whose functions in RNA are modulated in part by its reader proteins, we reason that environmental exposure-mediated changes in RNA pseudouridylation and the resulting changes in RNA-protein interactions constitute a major regulatory mechanism of RNA processes that may underlie disease pathogenesis. Yet up to now, few Ψ reader proteins are known, and our knowledge is limited in the mechanisms through which environmental exposure- mediated aberrant RNA pseudouridylation and the ensuing perturbations of RNA-protein interactions contribute to the pathogenesis of neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). Our proposed study aims to bridge this knowledge gap by uncovering the mechanistic link between environmental exposure and ALS pathology through the lens of RNA pseudouridylation and Ψ-protein interactions. Our preliminary data showed that: (1) at physiological concentration, profilin-1 (PFN1) binds directly and preferentially to Ψ-containing RNA; (2) ALS-linked point mutations in an ALS-associated protein, i.e., PFN1 reduce its affinity for Ψ-containing RNA, and (3) Ψ modification reduces RNA affinity for another ALS-associated protein. We organize our proposed research into three specific aims: Aim #1, to examine how RNA pseudouridylation is modulated by xenobiotic exposure in human neuronal cells; Aim #2, to investigate the impacts of RNA pseudouridylation on the transcriptome-wide occupancy of an ALS-associated protein; and Aim #3, to explore the impacts of RNA pseudouridylation and Ψ-protein interactions in xenobiotic and ALS pathological contexts. To accomplish these research aims and facilitate my transition to an independent academic career, I have assembled a mentoring committee with outstanding expertise in environmental toxicology, bioinformatics and neurobiology. The proposed research is built upon my extensive research skill sets, strong preliminary data, and the proposed mentoring/training plan. The outcome of the proposed research will unveil how environmental exposure-elicited aberrant RNA pseudouridylation alters RNA-protein interactions and contributes to ALS, thereby potentially leading to new biomarkers and therapeutic interventions. Moreover, the proposed career development and research plans will bridge gaps in my training and collect robust data for my independent publications and research grant applications, thereby transitioning me to an independent career at the intersection of environmental health science and neurodegeneration. Project Number: 1K99ES037399-01A1 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of Environmental Health Sciences (NIEHS) | Principal Investigator: Songbo Wei | Institution: UNIVERSITY OF CALIFORNIA RIVERSIDE, RIVERSIDE, CA | Award Amount: $120,663 | Activity Code: K99 | Study Section: Special Emphasis Panel[ZRG1 F12C-C (20)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11302126