Modeling Human Digestive Diseases With CRISPR-Cas9–Modified Organoids

Fujii, Masayuki; Clevers, Hans; Sato, Toshiro

doi:10.1053/j.gastro.2018.11.048

Cited by 115 publications

(79 citation statements)

References 152 publications

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“…The capacity to generate these preclinical models from surgical specimens and biopsies also has great potential to foster personalized medicine approaches to combine the highthroughput drug screening with the molecular assessment of organoid models in order to identify key diagnostic and therapeutic biomarkers that will predict which patients are at a higher risk of progression 49 . Furthermore, based on studies that have shown the ability of CRISPR/Cas9mediated genome editing to modify driver genes in human pancreatic ductal organoids, opportunity exists to replicate the genetic mechanisms leading to pancreatic tumorigenesis [50][51][52] .…”

Section: Discussionmentioning

confidence: 99%

Establishing a Living Biobank of Patient-Derived Organoids of Intraductal Papillary Mucinous Neoplasms of the Pancreas

Beato

Reverón

Dezsi

et al. 2020

Preprint

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Pancreatic cancer (PaCa) is the third leading cause of cancer-related deaths in the United States. There is an unmet need to develop strategies to detect PaCa at an early, operable stage and prevent its progression. Intraductal papillary mucinous neoplasms (IPMNs) are cystic PaCa precursors that comprise nearly 50% of pancreatic cysts detected incidentally via cross-sectional imaging. Since IPMNs can progress from low- and moderate-grade dysplasia to high-grade dysplasia and invasion, the study of these lesions offers a prime opportunity to develop early detection and prevention strategies. Organoids are an ideal preclinical platform to study IPMNs, and the objective of the current investigation was to establish a living biobank of patient-derived organoids (PDO) from IPMNs. IPMN tumors and adjacent normal pancreatic tissues were successfully harvested from 15 patients with IPMNs undergoing pancreatic surgical resection at Moffitt Cancer Center & Research Institute (Tampa, FL) between May of 2017 and March of 2019. Organoid cultures were also generated from cryopreserved tissues. Organoid count and size were determined over time by both Image-Pro Premier 3D Version 9.1 digital platform and Matlab application of a Circular Hough Transform algorithm, and histologic and genomic characterization of a subset of the organoids was performed using immunohistochemistry and targeted sequencing, respectively. The success rates for organoid generation from IPMN tumor and adjacent normal pancreatic tissues were 81% and 87%, respectively. IPMN organoids derived from different epithelial subtypes showed different morphologies in vitro, and organoids recapitulated histologic and genomic characteristics of the parental IPMN tumor. In summary, this pre-clinical model has the potential to provide new opportunities to unveil mechanisms of IPMN progression to invasion and to shed insight into novel biomarkers for early detection and targets for chemoprevention.

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Section: Discussionmentioning

confidence: 99%

Establishing a Living Biobank of Patient-Derived Organoids of Intraductal Papillary Mucinous Neoplasms of the Pancreas

Beato

Reverón

Dezsi

et al. 2020

Preprint

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“…in vivo orthotropic imaging, introduction of resistance, target validation, etc. A number of methods have proven useful in delivering gene editing into organoids, electroporation, lentiviral transduction and CRISPR [53][54][55] . Furthermore, genome-wide screens using RNAi-or sgRNA libraries 56 could be powerful tools for discovery of new drug targets.…”

Section: Discussionmentioning

confidence: 99%

A living biobank of matched pairs of patient-derived xenografts and organoids for cancer pharmacology

Xu¹,

Wang²,

Zhou³

et al. 2020

Preprint

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Patient-derived tumor xenograft (PDX)/organoid (PDO), driven by cancer stem cells (CSC), are considered the most predictive models for translational oncology. Large PDX collections reflective of patient populations have been created and used extensively to test various investigational therapies, including in population-trials as surrogate subjects in vivo. PDOs are recognized as in vitro surrogates for patients amenable for high-throughput screening (HTS). We have built a biobank of carcinoma PDX-derived organoids (PDXOs) by converting an existing PDX library and confirmed high degree of similarities between PDXOs and parental PDXs in genomics, histopathology and pharmacology, suggesting “biological equivalence or interchangeability” between the two. Here we demonstrate the applications of PDXO biobank for HTS “matrix” screening for both lead compounds and indications, immune cell co-cultures for immune-therapies and engineering enables in vitro/in vivo imaging. This large biobank of matched pairs of PDXs/PDXOs across different cancers could become powerful tools for the future cancer drug discovery.

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“…Complimentary approaches to the discovery process are also necessary given the mouse‐intensive nature of most current strategies and the fact that important human–microbiota interactions may not be modeled in a murine system. For example, primary intestinal organoids represent a particularly useful tool to study human epithelium–microbe interactions for several reasons, including: they can be generated directly from humans (with or without disease‐associated genetic polymorphisms); they can be genetically modified using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas technology; they recapitulate important epithelial cell lineages and physical features of the intestinal tract; and they can be seeded with microbes of interest . Another promising approach is to identify factors like metabolites that can impact a response of interest and work to identify the microbes that are responsible for their production .…”

Section: Of Mice and Menmentioning

confidence: 99%

“…For example, primary intestinal organoids represent a particularly useful tool to study human epithelium-microbe interactions for several reasons, including: they can be generated directly from humans (with or without disease-associated genetic polymorphisms); they can be genetically modified using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas technology; they recapitulate important epithelial cell lineages and physical features of the intestinal tract; and they can be seeded with microbes of interest. 93 Another promising approach is to identify factors like metabolites that can impact a response of interest and work to identify the microbes that are responsible for their production. 94,95 As knowledge of the key mechanisms of host-microbiota interactions are revealed, our accuracy in predicting immunomodulatory activity based on the genomic content of a strain or community should markedly improve.…”

Section: Iga-based Purificationmentioning

confidence: 99%

Understanding immune–microbiota interactions in the intestine

Ahern

Maloy

2019

Immunology

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Summary The past two decades have seen an explosion in research that aims to understand how the dynamic interplay with the gut microbiota impacts host health and disease, establishing a role for the gut microbiota in a plethora of pathologies. Understanding how health‐promoting microbiota are established and how beneficial host–microbiota interactions are maintained is of immense biomedical importance. Despite the enormous progress that has been made, our knowledge of the specific microbiota members that mediate these effects and the mechanisms underlying these interactions is rudimentary. The dearth of information regarding the nature of advantageous host–microbiota interactions, and the factors that cause these relationships to go awry, has hampered our ability to realize the therapeutic potential of the microbiota. Here we discuss key issues that limit current knowledge and describe a path forwards to improving our understanding of the contributions of the microbiota to host health.

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Modeling Human Digestive Diseases With CRISPR-Cas9–Modified Organoids

Cited by 115 publications

References 152 publications

Establishing a Living Biobank of Patient-Derived Organoids of Intraductal Papillary Mucinous Neoplasms of the Pancreas

Establishing a Living Biobank of Patient-Derived Organoids of Intraductal Papillary Mucinous Neoplasms of the Pancreas

A living biobank of matched pairs of patient-derived xenografts and organoids for cancer pharmacology

Understanding immune–microbiota interactions in the intestine

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