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Dissecting signalling pathways in prostate cancer using a unique primary prostate organoid culture system

Projectomschrijving

Prostaat- en darmkanker behoren tot de meest voorkomende en dodelijkste vormen van kanker. Onderzoek naar het ontstaan van deze vormen van kanker (en kanker in het algemeen) wordt ernstig belemmerd door het ontbreken van goede preklinische modelsystemen. Dit is één van de belangrijkste oorzaken van het falen van nieuw ontwikkelde medicijnen in klinische studies in patiënten. De recente ontwikkeling van de organoïdentechnologie maakt het mogelijk patiëntmateriaal in een kweekschaaltje te expanderen, met daarbij behoud van de fenotypische en genotypische kenmerken van het originele weefsel. In dit onderzoek hebben we organoïdenmodellen ontwikkeld voor gezond en kankerweefsel afkomstig van patiënten met prostaatkanker. Deze modellen kunnen vervolgens worden gebruikt voor het bestuderen van prostaatkanker en de ontwikkeling van nieuwe therapieën. Daarnaast hebben we gezonde dikke darmorganoïden gebruikt om colorectale tumoren te modelleren door kunstmatig genetische veranderingen (mutaties) te introduceren die veel voorkomen in dit type kanker. Op deze manier hebben we
darmkankermodellen ontwikkeld en deze gebruikt om te onderzoeken welke mutaties verantwoordelijk zijn voor het ontstaan van genetische instabiliteit (een belangrijk kenmerk van tumorcellen), en metastasering van tumorcellen.  Dit unieke dikke darmkankerprogressie model stelt ons in staat om de mutaties te bestuderen welke ten grondslag liggen aan
het ontstaan van kanker. Onze data suggereren dat slechts vier mutaties al voldoende zijn om een gezonde cel te transformeren in een metastaserende tumorcel. Onze bevindingen bieden mogelijk nieuwe targets voor therapeutische interventie. 

We laten zien dat onze modellen de patiëntsituatie beter nabootsen dan andere tot nu toe gebruikte modelsystemen. De bevindingen in dit project hebben nieuwe deuren geopend voor het bestuderen van de genetische veranderingen die ten grondslag liggen aan het ontstaan van kanker en de ontwikkeling van nieuwe therapieën.

Producten

Titel: Translational applications of adult stem cell-derived organoids
Auteur: Drost, Jarno, Clevers, Hans
Magazine: Development
Titel: Identification of Multipotent Luminal Progenitor Cells in Human Prostate Organoid Cultures
Auteur: Karthaus, Wouter R., Iaquinta, Phillip J., Drost, Jarno, Gracanin, Ana, van Boxtel, Ruben, Wongvipat, John, Dowling, Catherine M., Gao, Dong, Begthel, Harry, Sachs, Norman, Vries, Robert G.J., Cuppen, Edwin, Chen, Yu, Sawyers, Charles L., Clevers, Hans C.
Magazine: Cell
Titel: TGFß signaling directs serrated adenomas to the mesenchymal colorectal cancer subtype.
Auteur: Evelyn Fessler, Jarno Drost, Sander R van Hooff, Janneke F Linnekamp, Xin Wang, Marnix Jansen, Felipe De Sousa E Melo, Pramudita R Prasetyanti, Joep EG IJspeert, Marek Franitza, Peter Nürnberg, Carel JM van Noesel, Evelien Dekker, Louis Vermeulen, Hans Clevers, Jan Paul Medema
Magazine: EMBO Molecular Medicine
Titel: Genetic dissection of colorectal cancer progression by orthotopic transplantation of engineered cancer organoids
Auteur: Fumagalli, Arianna, Drost, Jarno, Suijkerbuijk, Saskia J. E., van Boxtel, Ruben, de Ligt, Joep, Offerhaus, G. Johan, Begthel, Harry, Beerling, Evelyne, Tan, Ee Hong, Sansom, Owen J., Cuppen, Edwin, Clevers, Hans, van Rheenen, Jacco
Magazine: Proceedings of the National Academy of Sciences USA
Titel: Who Is in the Driver’s Seat: Tracing Cancer Genes Using CRISPR-Barcoding
Auteur: Drost, Jarno, Clevers, Hans
Magazine: Molecular Cell
Titel: Use of CRISPR-modified human stem cell organoids to study the origin of mutational signatures in cancer
Auteur: Drost, Jarno, van Boxtel, Ruben, Blokzijl, Francis, Mizutani, Tomohiro, Sasaki, Nobuo, Sasselli, Valentina, de Ligt, Joep, Behjati, Sam, Grolleman, Judith E., van Wezel, Tom, Nik-Zainal, Serena, Kuiper, Roland P., Cuppen, Edwin, Clevers, Hans
Magazine: Science
Titel: Sequential cancer mutations in cultured human intestinal stem cells
Auteur: Drost, Jarno, van Jaarsveld, Richard H., Ponsioen, Bas, Zimberlin, Cheryl, van Boxtel, Ruben, Buijs, Arjan, Sachs, Norman, Overmeer, René M., Offerhaus, G. Johan, Begthel, Harry, Korving, Jeroen, van de Wetering, Marc, Schwank, Gerald, Logtenberg, Meike, Cuppen, Edwin, Snippert, Hugo J., Medema, Jan Paul, Kops, Geert J. P. L., Clevers, Hans
Magazine: Nature
Titel: Targeting mutant RAS in patient-derived colorectal cancer organoids by combinatorial drug screening
Auteur: Verissimo, Carla S, Overmeer, René M, Ponsioen, Bas, Drost, Jarno, Mertens, Sander, Verlaan-Klink, Ingrid, Gerwen, Bastiaan van, van der Ven, Marieke, Wetering, Marc van de, Egan, David A, Bernards, René, Clevers, Hans, Bos, Johannes L, Snippert, Hugo J
Magazine: eLife
Titel: Organoid culture systems for prostate epithelial and cancer tissue
Auteur: Drost, Jarno, Karthaus, Wouter R, Gao, Dong, Driehuis, Else, Sawyers, Charles L, Chen, Yu, Clevers, Hans
Magazine: Nature Protocols
Titel: The Generation of Organoids for Studying Wnt Signaling.
Auteur: Jarno Drost, Benedetta Artegiani, Hans Clevers

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Samenvatting van de aanvraag

Prostate cancer is worldwide the second most common type of cancer in men. Although many advances were made over the last decade in the early detection and treatment of localized prostate cancer, it is still the second most common cause of male cancer death, mainly caused by metastatic growth. Androgens are essential for normal prostate development and homeostasis and androgen deprivation is therefore currently the main treatment therapy. However, treatment resistance occurs in the majority of the cases. Additionally, the current prostate cancer screening, based on elevated blood prostate-specific antigen (PSA) levels and subsequent prostate biopsy, has a major disadvantage: it cannot distinguish indolent from aggressive tumours. This regularly leads to overtreatment of indolent tumours, which also accounts for a significant amount of deaths. Therefore, there is a need for biomarkers that could predict disease outcome and for novel therapy targets. Most research on prostate cancer is currently performed using prostate cancer cell lines. These are few in number, were derived from metastatic lesions, and their establishment as a cell line required adaptations to 2D culture conditions. The accumulation of in vitro generated genetic aberrations renders them poor representatives of in vivo tumorigenesis. I will make use of a newly developed unique three-dimensional culture system for human and mouse primary prostate tissue (organoids). These prostate organoids have gland-like structures, consist of the different prostate epithelial cell lineages (basal and luminal) and are dependent for their growth on the androgen testosterone. The innovative organoid technology allows me to, for the first time, study signalling pathways and genetics involved in prostate development and cancer in vitro. With this system I will study the regulation of normal development and tumorigenesis using state of the art techniques such as mass spectrometry, RNA expression analysis combined with conventional biochemistry and cell biology. I will perform this research in the lab of Dr. Clevers, world leader in the field of stem cell and cancer biology and I will make use of already existing collaborations with leaders in the field of prostate cancer research, mass spectrometry and cancer genome sequencing. First, I will use this model system to characterize genes and pathways that are altered in primary and treatment-resistant prostate cancer and of which the functional contribution is unknown. I will functionally and mechanistically study these genes using RNA interference and gene overexpression in prostate organoids and determine the effect on proliferation and differentiation. Second, I will investigate the contribution of different signalling pathways in prostate epithelial cell lineage differentiation. The molecular pathways involved in basal and luminal cell differentiation (the two main epithelial cell lineages of the prostate) are largely unknown. Although prostate tumours have a luminal phenotype, it remains unclear whether the luminal lineage is indeed the source of the tumour-initiating cell, or that it originates from basal cells that lose their characteristics during tumorigenesis. Therefore, additional research is required to unravel prostate lineage development. The prostate organoids are cultured in defined conditioned medium, which allows the study of individual signalling pathways. I will determine the contribution of the Wnt, Notch, TGFβ and BMP pathways (all known to be involved in lineage specification in other organs, including small intestine) to prostate lineage differentiation. Finally, a biobank will be generated containing organoids derived from normal and tumour tissue of prostate cancer patients. The extremely high success rate of the establishment of patient-derived organoids enables me to directly compare genome sequence data, RNA expression data with in vitro biochemistry and cell biology in parallel in patients. In collaboration with the Welcome Trust Sanger Institute (UK), we will also perform drug screens on normal and tumour organoids to find new compounds that specifically inhibit growth of tumour organoids. Currently, drug screening relies on cell lines and (xenograft) animal models with their limitations. The organoid culture system provide an unlimited source of patient-derived, genetically, and clinically stable material for drug screening and validation. Altogether, the research I propose here will add to the understanding of the underlying mechanisms in normal prostate development and prostate cancer initiation and progression. Identified genes might ultimately serve as novel biomarkers or targets for prostate cancer therapy.

Onderwerpen

Kenmerken

Projectnummer:
91614138
Looptijd: 100%
Looptijd: 100 %
2014
2017
Gerelateerde subsidieronde:
Projectleider en penvoerder:
Prof. dr. H.C. Clevers
Verantwoordelijke organisatie:
Hubrecht Institute