Investigating cellular adaptation to one-carbon metabolism perturbation

Cells respond to nutrient availability and adjust their functions and performance to ensure survival. This adaptation requires rapid, nutrient-responsive decision-making and prioritization of essential cellular processes. For example, we recently showed that depletion of the essential vitamin folate in erythroid cells results in premature differentiation that improves cell survival on the expense of proliferation rate. Our work, that showed that erythroid cell lines as well as primary murine erythroid progenitors prematurely differentiate under folate deficiency, revealed a potential etiology for folate deficiency-induced anemia. To further reveal the cellular response to folate deprivation and other perturbations of the one-carbon (1C) metabolism pathway in erythroid cells, we performed a genome wide CRISPR screen and RNAseq in folate deprived erythroid cells. Our CRISPR screen revealed all five genes of the post translational modification (PTM) UFMylation pathway as top hits, and the RNAseq data showed differential gene expression induction following folate deprivation through key signaling pathways including mTOR. Although recent literature defines the crucial role of UFMylation in mediating ER stress and homeostasis of translation, the specific triggers and consequences of UFMylation remain largely unknown, warranting further exploration of conditions which trigger this PTM. We hypothesize that regulation of UFMylation maintains cellular fitness during folate depletion. To address this hypothesis, we will validate our CRISPR screen results, characterize the role of UFMylation in cellular adaptation to 1C disruption, identify UFMylation targets, and explore the role of UFMylation in nutritional status-dependent ribosomal regulation. We further hypothesize that 1C perturbation induces a signaling cascade to promote cellular fitness. To address this hypothesis, we will pharmacologically and genetically target pathways known to be differentially activated during nutrient sensing and probe the role of these pathways in cell function regulation during 1C metabolism disruption. Completing these aims will mark a new role of UFMylation in a signaling pathway activated in response to metabolic perturbation and define a new cellular adaptation mechanism in response to 1C metabolism disruption. As a Ph.D. student in the Kanarek lab I have the commitment and support of my sponsors Dr. Kanarek and Dr. Toker, as well as access to the expertise of leaders in the metabolism and cell signaling fields, and to state-of-the-art facilities and equipment. Leveraging these resources, I plan to achieve the following over the next three years: (1) Gain proficiency in molecular and cellular biology techniques for rigorous scientific research in metabolism; (2) Deepen my knowledge of nutrient sensing and erythroid biology, contributing to basic biomedical research; (3) Enhance communication skills through writing, presenting, and networking to build a mentorship and peer network to support my future in biomedical research; (4) Develop leadership skills through mentoring students at various academic stages, drawing on my own experiences. Project Number: 1F31HL178310-01 | Fiscal Year: 2025 | NIH Institute/Center: National Heart Lung and Blood Institute (NHLBI) | Principal Investigator: Nancy Pohl | Institution: BOSTON CHILDREN'S HOSPITAL, BOSTON, MA | Award Amount: $39,551 | Activity Code: F31 | Study Section: Special Emphasis Panel[ZRG1 F05-Q (20)] View on NIH RePORTER: https://reporter.nih.gov/project-details/1F31HL17831001