Abstract
Abstract Cells are compartmentalized into “classical” membrane-bound organelles such as the endoplasmic reticulum and mitochondria. Cells also have membrane-less organelles (MLOs), which are formed through phase separation. Examples of MLOs include stress granules and nucleoli, where macromolecules (proteins, DNAs, RNAs) coherently and dynamically concentrate into biomolecular condensates resembling droplets in lava lamps. MLOs play central role in cellular processes - choreographed events such as transcription and RNA splicing take place in MLOs. MLOs are highly organized, and their dysregulation always led to diseases. Papillary renal cell carcinoma (PRCC) is the second most common RCC in America. PRCC is an aggressive cancer, yet the uncharacterized molecular etiology results in the lack of effective treatment for patients. Our lab and others have observed dysregulated oncoproteins MLOs in PRCC. However, the molecular properties and functions of these aberrant MLOs have not been characterized. The photolyase homology region of Arabidopsis thaliana Cry2 (Cry2WT) is a light-sensitive protein that self-associate with blue light exposure. We hypothesize that fusing dysregulated MLOs oncoproteins with Cry2WT will allow us to tune and study the oncoproteins condensates. We aim to develop an optogenetic model to study how oncoproteins condensates function in PRCC cells. Dysregulated oncoproteins were fused to mCherry-Cry2WT, introduced into cell line models, light-activated and imaged using a ZEISS LSM 900 microscope. We found that blue-light indeed induced proteins clustering when oncoproteins were fused to mCherry-Cry2WT but not with mCherry-Cry2WT alone. This model will allow us to tune the formation of oncoproteins condensates. Future work will involve studying how optogenetically tuned oncoproteins condensates function in PRCC patient-derived cell lines. Citation Format: Choon Leng So, Jane Lee, Binglin Huang, Danfeng Cai. Developing a model to investigate the roles of biomolecular condensates in papillary renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1495.
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