Heterogeneous and novel transcript expression in single cells of patient-derived clear cell renal cell carcinoma organoids

Splicing is often dysregulated in cancer, leading to alterations in the expression of canonical and alternatively spliced isoforms. We used the multiplexed arrays sequencing (MAS-seq) protocol of PacBio to sequence full-length transcripts in patient-derived organoid (PDO) cells of clear cell renal cell carcinoma (ccRCC). The sequencing revealed a heterogeneous dysregulation of splicing across 2599 single ccRCC cells. The majority of novel transcripts could be removed with stringent filtering criteria. The remaining 31,531 transcripts (36.6% of the 86,182 detected transcripts on average) were previously uncharacterized. In contrast to known transcripts, many of the novel transcripts have cell-specific expression. Novel transcripts common to ccRCC cells belong to genes involved in ccRCC-related pathways, such as hypoxia and oxidative phosphorylation. A novel transcript of the ccRCC-related gene nicotinamide N-methyltransferase is validated using PCR. Moreover, >50% of novel transcripts possess a predicted complete protein-coding open reading frame. An analysis of the most dominant transcript-switching events between ccRCC and non-ccRCC cells shows many switching events that are cell- and sample-specific, underscoring the heterogeneity of alternative splicing events in ccRCC. Overall, our study elucidates the intricate transcriptomic architecture of ccRCC, underlying its aggressive phenotype and providing insights into its molecular complexity.

Keywords

Alternative splicing, Carcinoma, Renal cell, Gene expression profiling, Gene expression regulation, Neoplastic, Humans, Kidney neoplasms, Organoids, Single-Cell analysis, Transcriptome

Subject (DDC)

610 - Medizin und Gesundheit

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ISBN

ISSN

1088-9051
1549-5469

Language

English

Created during FHNW affiliation

Yes

Strategic action fields FHNW

Publication status

Published

Review

Peer review of the complete publication

Open access category

Hybrid

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Citation

Karakulak, T., Zajac, N., Bolck, H. A., Bratus-Neuenschwander, A., Zhang, Q., Qi, W., Basu, D., Oltra, T. C., Rehrauer, H., von Mering, C., Moch, H., & Kahraman, A. (2025). Heterogeneous and novel transcript expression in single cells of patient-derived clear cell renal cell carcinoma organoids. Genome Research, 35(4), 698–711. https://doi.org/10.1101/gr.279345.124

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