Project ID CM-HD2026_32

ThemeCM-HD

Co Supervisor 1A Dr Rocio Sancho Faculty of Life Sciences & Medicine, School of Basic & Medical Biosciences, Randall Centre for Cell & Molecular BiophysicsEmail

Co Supervisor 1B Dr Alessandra Vigilante Faculty of Life Sciences & Medicine, School of Basic & Medical Biosciences, Randall Centre for Cell & Molecular BiophysicsEmail

Stem Cell-derived Pancreatic Organoids: elucidating molecular regulators of beta cell differentiation for regenerative diabetes therapies

Diabetes type 1 is caused by irreversible loss of insulin-producing beta cells in the pancreas. There are currently no cures, and most patients manage their disease with insulin injections. In the last decade, stem cell research has accelerated research for pancreatic replacement therapies. At this moment however, differentiation of stem cells into functional beta cells is still inefficient. In our lab, we use pancreatic organoids differentiated from skin-derived induced pluripotent stem cells (iPSCs) from human donors. This model allows us to study the molecular mechanisms underlying beta cell differentiation and has led to the finding of USP7 as a key player in beta cell differentiation. USP7 is a deubiquitinating enzyme that mediates degradation rate of specific proteins, including embryonic transcription factor NGN3. Its other roles in beta cell differentiation remain to be explored.

The goal of the proposed PhD project is to identify novel USP7 pathways that regulate beta cell differentiation.

• During the rotation project the PhD student will set up iPSC to organoid differentiation experiments. The student will use the labs iPSC cell lines with inducible Usp7-overexpression.
• The goal of year 1/2 is to perform single cell RNA-sequencing of pancreatic organoids after Usp7 over-expression, to analyse the transcriptomics data, and identify possible interactors.
• In year 2/3 the student will validate the top 10 hits in biochemical and functional assays. The student will have access to state-of-the-art facilities for fluorescent microscopy and flow cytometry to carry out these experiments.
• In the final year, the most promising candidates will be further analysed for physiological effects in the mature beta cell. This could range from in vitro assays (e.g. insulin release, calcium metabolism) to in vivo experiments (e.g. mouse kidney transplantation).

During this project, the PhD student will be part of our enthusiastic and friendly team here at the Centre for Gene Therapy and Regenerative Medicine. They will have access to the broad expertise of the centre and can gain from its collaborative spirit.

Representative Publications

1. Manea T, Nelson JK, Garrone CM, Hansson K, Evans I, Behrens A, and Sancho R. 2023. ‘USP7 Controls NGN3 Stability and Pancreatic Endocrine Lineage Development’. Nature Communications 14 (1): 2457. https://doi.org/10.1038/s41467-023-38146-9.
2. Pedraza-Arevalo S, Cujba AM, Alvarez-Fallas ME, and Sancho R. 2022. ‘Differentiation of Beta-like Cells from Human Induced Pluripotent Stem Cell-Derived Pancreatic Progenitor Organoids’. STAR Protocols 3 (3): 101656. https://doi.org/10.1016/j.xpro.2022.101656.
3. Cujba AM, Alvarez-Fallas ME, Pedraza-Arevalo S, Laddach A, Shepherd MH, Hattersley A, Watt FM and Sancho R. 2022. ‘An HNF1α truncation associated with maturity-onset diabetes of the young impairs pancreatic progenitor differentiation by antagonizing HNF1β function’. Cell Reports 38 (9): 110425. https://doi.org/10.1016/j.celrep.2022.110425

1. Willnow D, Benary U, Margineanu A, Vignola ML, Konrath F, Pongrac IM, Karimaddini Z, Vigilante A, Wolf J, Spagnoli FM. Quantitative lineage analysis identifies a hepato-pancreato-biliary progenitor niche. Nature. 2021 Sep;597(7874):87-91. doi: 10.1038/s41586-021-03844-1. Epub 2021 Aug 25. PMID: 34433966.
2. Vigilante A, Laddach A, Moens N, Meleckyte R, Leha A, Ghahramani A, Culley OJ, Kathuria A, Hurling C, Vickers A, Wiseman E, Tewary M, Zandstra PW; HipSci Consortium; Durbin R, Fraternali F, Stegle O, Birney E, Luscombe NM, Danovi D, Watt FM. Identifying Extrinsic versus Intrinsic Drivers of Variation in Cell Behavior in Human iPSC Lines from Healthy Donors. Cell Rep. 2019 Feb 19;26(8):2078-2087.e3. doi: 10.1016/j.celrep.2019.01.094. PMID: 30784590; PMCID: PMC6381787.
3. Segal JM, Kent D, Wesche DJ, Ng SS, Serra M, Oulès B, Kar G, Emerton G, Blackford SJI, Darmanis S, Miquel R, Luong TV, Yamamoto R, Bonham A, Jassem W, Heaton N, Vigilante A, King A, Sancho R, Teichmann S, Quake SR, Nakauchi H, Rashid ST. Single cell analysis of human foetal liver captures the transcriptional profile of hepatobiliary hybrid progenitors. Nat Commun. 2019 Jul 26;10(1):3350. doi: 10.1038/s41467-019-11266-x. PMID: 31350390; PMCID: PMC6659636.