Human organoids: model systems for human biology and medicine

Nature Reviews Molecular Cell Biology, Journal Year: 2020, Volume and Issue: 21(10), P. 571 - 584

Published: July 7, 2020

Language: Английский

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Engineering organoids

Nature Reviews Materials, Journal Year: 2021, Volume and Issue: 6(5), P. 402 - 420

Published: Feb. 19, 2021

Language: Английский

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A Patient-Derived Glioblastoma Organoid Model and Biobank Recapitulates Inter- and Intra-tumoral Heterogeneity

Cell, Journal Year: 2019, Volume and Issue: 180(1), P. 188 - 204.e22

Published: Dec. 26, 2019

Language: Английский

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Cancer modeling meets human organoid technology

Science, Journal Year: 2019, Volume and Issue: 364(6444), P. 952 - 955

Published: June 6, 2019

Organoids are microscopic self-organizing, three-dimensional structures that grown from stem cells in vitro. They recapitulate many structural and functional aspects of their vivo counterpart organs. This versatile technology has led to the development novel human cancer models. It is now possible create indefinitely expanding organoids starting tumor tissue individuals suffering a range carcinomas. Alternatively, CRISPR-based gene modification allows engineering organoid models through introduction any combination alterations normal organoids. When combined with immune fibroblasts, become for microenvironment enabling immune-oncology applications. Emerging evidence indicates can be used accurately predict drug responses personalized treatment setting. Here, we review current state future prospects rapidly evolving field.

Language: Английский

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Establishment of patient-derived cancer organoids for drug-screening applications

Nature Protocols, Journal Year: 2020, Volume and Issue: 15(10), P. 3380 - 3409

Published: Sept. 14, 2020

Language: Английский

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Organoids

Nature Reviews Methods Primers, Journal Year: 2022, Volume and Issue: 2(1)

Published: Dec. 1, 2022

Organoids have attracted increasing attention because they are simple tissue-engineered cell-based in vitro models that recapitulate many aspects of the complex structure and function corresponding vivo tissue. They can be dissected interrogated for fundamental mechanistic studies on development, regeneration, repair human tissues. also used diagnostics, disease modeling, drug discovery, personalized medicine. derived from either pluripotent or tissue-resident stem (embryonic adult) progenitor differentiated cells healthy diseased tissues, such as tumors. To date, numerous organoid engineering strategies support culture growth, proliferation, differentiation maturation been reported. This Primer serves to highlight rationale underlying selection development these materials methods control cellular/tissue niche; therefore, engineered organoid. We discuss key considerations generating robust organoids, those related cell isolation seeding, matrix soluble factor selection, physical cues integration. The general standards data quality, reproducibility deposition within community is outlined. Lastly, we conclude by elaborating limitations organoids different applications, priorities coming years.

Language: Английский

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A brief history of organoids

AJP Cell Physiology, Journal Year: 2020, Volume and Issue: 319(1), P. C151 - C165

Published: May 27, 2020

In vitro cell cultures are crucial research tools for modeling human development and diseases. Although the conventional monolayer have been widely used in past, lack of tissue architecture complexity such model fails to inform true biological processes vivo. Recent advances organoid technology revolutionized culture biomedical by creating powerful three-dimensional (3D) models recapitulate cellular heterogeneity, structure, functions primary tissues. Such enables researchers recreate organs diseases a dish thus holds great promises many translational applications as regenerative medicine, drug discovery, precision medicine. this review, we provide an overview history development. We discuss strengths limitations organoids well their potential laboratory clinic.

Language: Английский

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Single-Cell Atlas of Lineage States, Tumor Microenvironment, and Subtype-Specific Expression Programs in Gastric Cancer

Cancer Discovery, Journal Year: 2021, Volume and Issue: 12(3), P. 670 - 691

Published: Oct. 12, 2021

Gastric cancer heterogeneity represents a barrier to disease management. We generated comprehensive single-cell atlas of gastric (>200,000 cells) comprising 48 samples from 31 patients across clinical stages and histologic subtypes. identified 34 distinct cell-lineage states including novel rare cell populations. Many lineage exhibited cancer-associated expression profiles, individually contributing combined tumor-wide molecular collage. observed increased plasma proportions in diffuse-type tumors associated with epithelial-resident KLF2 stage-wise accrual fibroblast subpopulations marked by high INHBA FAP coexpression. Single-cell comparisons between patient-derived organoids (PDO) primary highlighted inter- intralineage similarities differences, demarcating boundaries PDOs as experimental models. complemented these findings spatial transcriptomics, orthogonal validation independent bulk RNA-sequencing cohorts, functional demonstration using vitro vivo Our results provide high-resolution resource intra- interpatient profiled malignancies at resolution feature tumors. also uncovered subtypes INHBA-FAP-high populations predictors poor prognosis. highlight potential origins deregulated the tumor ecosystem. This article is In Issue feature, p. 587.

Language: Английский

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Xenograft and organoid model systems in cancer research

The EMBO Journal, Journal Year: 2019, Volume and Issue: 38(15)

Published: July 8, 2019

Review8 July 2019Open Access Xenograft and organoid model systems in cancer research Margit Bleijs Oncode Institute, Princess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands Search more papers by this author Marc van de Wetering Hans Clevers orcid.org/0000-0002-3077-5582 Hubrecht Royal Academy of Arts Sciences University Medical Center, Jarno Drost Corresponding Author [email protected] orcid.org/0000-0002-2941-6179 Information Bleijs1, Wetering1, Clevers1,2 *,1 1Oncode 2Oncode *Corresponding author. Tel: +31 88 972 72 72; E-mail: EMBO Journal (2019)38:e101654https://doi.org/10.15252/embj.2019101654 PDFDownload PDF article text main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Patient-derived tumour xenografts organoids have become important preclinical research. Both models maintain key features from their parental tumours, such as genetic phenotypic heterogeneity, which allows them be used a wide spectrum applications. In contrast patient-derived xenografts, can established expanded with high efficiency primary patient material. On the other hand, retain tumour–stroma interactions, are known contribute tumorigenesis. review, we discuss recent advances xenograft compare promises challenges Introduction Cancers consist continuously evolving heterogeneous cell mass (McGranahan Swanton, 2017). Importantly, not all cells within equally its progression. Early studies on mouse mammary tumours revealed that cellular subpopulations different regions same vary growth rate, drug response, immunogenicity metastatic capacity (reviewed Heppner, 1984; Tabassum Polyak, 2015). This intra-tumour heterogeneity arise both non-genetic variability variations availability resources, like differential access oxygen nutrients (Kreso Dick, 2014; development phenocopying is required studying contribution progression acquisition therapy resistance. Whereas first (PDTX) were successfully during fifties (Toolan, 1953), (PDTO) been only last decade (Sato et al, 2011) (Fig 1). PDTX PDTO able recapitulate intra- inter-tumour seen human cancers (Beckhove 2003; Guenot 2006; Huang 2015; Bruna 2016; George 2017; Nanki 2018; Sachs Yan 2018). Therefore, these promising tools study sub-clonal dynamics individual resistance (Shi 2014). Figure 1. Timeline development(Toolan, 1953; Taetle 1987; Fu 1992; Beckhove Fichtner 2004; Shultz 2005; Wang Cutz Sato 2009, 2011; Hennessey Gao Karthaus Boj Lee Li 2018, 2019; Kopper Schutgens 2019). Download figure PowerPoint Due complexity response clinical treatments varies substantially. Additionally, mechanisms poorly defined well efficacy While number anti-cancer compounds tested safety Phase I progress II testing, most fail III studies, examine power pharmacological responses (Dimasi 2013). Such failure rates trials headline need improved predictions outcome. Several currently used, including lines, cultures (Figs 1 2). These our understanding provided valuable novel treatments. predict agents. 2. Schematic representation organoidsPDTXs preserve interactions. PDTOs grow basement membrane extract epithelial normal tissue. allow several translational applications therapeutic Part was adapted (2014). To understand biology translation into effective treatments, capturing fundamental. Although low efficiency, tissues grown 2D vitro, allowing capable adapting conditions expand form line. use vitro lines has insights actions (Sos 2009; Greshock 2010). However, drawback lack found original (Sachs Clevers, Byrne enhance correlation cancers, surgically derived samples grafted mice, PDTX. models, architecture relative proportion stromal maintained large extent, yields better resemblance compared (Byrne Fig engrafted success rate establishment increasing due immunocompromised mice (Shultz Drake 2012). documented attempt transplant an animal dates back 1775, hallmark Helene Toolan showed it possible x-irradiated rats 1953). she demonstrated proliferation extended considerably when x-radiated hosts treated immune system suppressor cortisone Later, Phillips Gazet obtain slightly higher percentage viable grafts treating host anti-lymphocyte serum. increased particular combined thymectomy, demonstrating suppressed enhances engraftment (Phillips Gazet, 1970). Following genetically modified established, severely deficient, NOD/SCID/IL2Rγnull (NSG) 2005). virtual absence significantly Together loss activity improves viability tissue mice. broad variety colorectal (Fichtner 2006), pancreatic (Fu Kim 2009), breast 2016), lung (Cutz skin (Taetle 1987), head neck (Hennessey 2011), prostate (Wang 2005) ovarian (George 3). PDTXs closely than they usually generated small amount As consequence, might capture full (Kemper Moreover, Morgan colleagues recently reported total mutations detected non-small-cell-lung (NSCLC) 43% corresponding four additional arose early passages present (Morgan observations suggest clonal selection evolution may occur upon Multiple biopsies should complete vivo, whereas avoid outcomes deviate response. Furthermore, limited remain major challenge, highly variable among (Rosfjord Sub-clones advanced best PDTX, less tumours. increase over significant between passage grade (Pearson 2016). indicates occurs PDTXs. 3. Pie chart types (left) (right) marked greenIn general, lower establishment. stroma remains controversial models. Components stroma, vasculature, fibroblasts, presence components interaction microenvironment. subsequently replaced murine (Julien 2012; Peng Gene expression NSCLC confirmed depletion human-derived tumour-associated downregulation genes adhesion pathways. suggests deviates time addition, metabolism pharmacokinetics differ human, needs taken account Over years, increasingly clear orthotopic transplantation provide physiological heterotopic (e.g. subcutaneous) engraftment. It previously lead local invasive metastases, similar those observed patients (Dai Hoffman, PDTXs, tumour-host interactions investigated at relevant location secondary growth, metastases. comparison subcutaneous ductal adenocarcinoma (PDAC), metabolic differences found. could attributed microenvironment caused (Zhan results highlight importance environment site. Nonetheless, while accurately mimic native microenvironment, method technically challenging labour-intensive. still During decade, techniques 3D organotypic structures established. so-called adult embryonic stem self-organize reflect origin (for cell-derived organoids), or differentiation directed (embryonic organoids) review see Lgr5-expressing intestinal placed mimicking niche 2009). By providing R-spondin-1, epidermal factor (EGF) Noggin, embedment extracellular matrix-providing membranes extract, received signalling cues necessary self-renew, proliferate differentiated offspring, resembling epithelium Since then, tissues, (Hild Jaffe, Tan 2019), colon stomach (Bartfeld 2015), liver (Huch pancreas (Boj (Chua 2014), kidney (Jun 2019) fallopian tube (Kessler culture protocols well. Human (Gao 2018), gastric (Nanki oesophageal (Li bladder (Lee Mullenders (Kopper (Schutgens (Broutier An feature phenotypically mirror epithelium, (Huang study, Roerink characterized single (CRC) extensive mutational diversification drugs even related (Roerink show cultured lines. Thereby, bridge gap line vivo long term cryopreserved, generation living biobanks (Weeber Fujii Schütte Seino Tiriac So far, majority originate (carcinomas). common carcinomas origin, not, sarcomas, leukaemia lymphomas. challenge technology cancers. cannot vasculature tool applications, high-throughput screens personalized medicine manner. Translational Most agents entering acquire regulatory approval insufficient inefficacy. highlights limitations predictive value current where evaluated relevance panel six small-cell-lung carcinoma (SCLC) topotecan, topoisomerase inhibitor, combinations topotecan inhibitor etoposide alkylating ifosfamide cisplatin maximum tolerated dose (Némati Three out 90% inhibition alone, (Ardizzoni 1997). Growth ifosfamide. findings demonstrate useful assessment new 2010; Rosfjord Bertotti screened cohort 85 CRC (mCRC) cetuximab, inhibiting antibody against receptor (EGFR). They enrichment HER2 amplification cetuximab-resistant KRAS/NRAS/BRAF/PIK3CA wild-type (Bertotti 2011). proof-of-concept EGFR induced long-lasting regression, suggesting opportunities mCRC resistant cetuximab collection 1,000 representing range solid hypotheses biomarkers sensitivity validated. also identified candidates failed testing unravel For example, metformin, anti-diabetic drug, affects Administering metformin levels 150 mg/kg per day equivalent 500–1,000 mg/daily sufficient inhibit implies options (Suhaimi Together, develop patients. Fast expansion enable screens. easily detection gene–drug associations Verissimo KRAS pathway inhibitors harbouring (Verissimo another Vlachogiannis biobank metastatic, heavily pretreated gastroesophageal degree similarity A PDTO-based trials, (Vlachogiannis library largely transcriptional subtypes cancer. exhibited standard-of-care chemotherapeutics profiles correspond outcomes. data molecular profiling treatment prospective (Tiriac collected 106 CRCs, 35 59 identify linking patterns. landscape maintained, some PDTOs, appeared closer distinct groups PDTOs. elevated involved xenobiotic fatty acid processes, affect (Schütte Organoid additionally engineering effects oncogenic detail. introduced colon, pancreas. shows presented dysplasia result activating KrasG12D mutation, Tp53 formed transplantation. contrast, combinatorial Apc, Tp53, Smad4 formation vivo. Opposed organoids, intestine rapid combination mutated Apc Kras Subsequently, two translated situation CRISPR/Cas9-mediated genome editing driver healthy colonic (Drost Matano mutation KRAS, inactivating APC, TP53 SMAD4, independent factors EGF, Wnt, R-spondin Noggin. al APC drivers chromosome instability aneuploidy Not initiation progression, but towards progenitor acinar structures. Expression mutant mutation-specific phenotypes. instance, TP53, cytosolic SOX9 localization, associated mortality CRISPR-modified DNA repair defects accumulation deficient mismatch gene MLH1 repair-deficient CRC. Application approach predisposition NTHL1 footprint (signature 30), cohort, indicative germline immense gives specific alterations Integrating resides, plays role development. Stromal modulate behaviour directly influence (Tauriello effectively shown Batlle colleagues, using specifically cells. Quadruple-mutant developed hallmarks CRC, T-cell exclusion TGFβ-activated stroma. TGFβ cytotoxic prevented metastasis, highlighting Immune recognize antigens distinguish non-cancer (Schumacher Schreiber, consequence tumour-specific mutations, start expressing neoantigens membranes, recognized T lymphocytes. recognition immunotherapies. neoantigen load biomarker competent immune-deficient limits utility explore system. overcome limitation humanized (Box this, selected establish (HIS) Humanized various lineages blood throughout lifetime recipient animal. Ideally, hematopoietic (HSCs) come whom will order reactions leucocyte antigen mismatch. challenging, because bone marrow burden weakened factor-stimulated mobilization collecting HSCs peripheral support (Voloshin CD34-positive obtainable strongly limit anti-tumour newborn NSG co-engrafted accompanied maturation cell-specific activation NK site (Wege Transplantation existence tumour-infiltrating effector memory activated IL-12 administration (Simpson-Abelson 2008). ability provides approaches immunotherapy, therapies involvement chemotherapy. cultures, co-culture Nozaki intra-epithelial lymphocytes co-culture, IL-2, IL-7 IL-15 (Nozaki Recently, Zumwalde (2016) succeeded characterizing lymphocyte compartment identifying subset pharmacologically targeted Specifically, Vδ2+ γδ constantly preparation organoids. zoledronic acid, aminobisphosphonate started proliferate.

Language: Английский

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Disease modelling in human organoids

Disease Models & Mechanisms, Journal Year: 2019, Volume and Issue: 12(7)

Published: July 1, 2019

ABSTRACT The past decade has seen an explosion in the field of vitro disease modelling, particular development organoids. These self-organizing tissues derived from stem cells provide a unique system to examine mechanisms ranging organ homeostasis and disease. Because organoids develop according intrinsic developmental programmes, resultant tissue morphology recapitulates architecture with remarkable fidelity. Furthermore, fact that these can be human progenitors allows for study uniquely processes disorders. This article accompanying poster highlight currently available methods, particularly those aimed at modelling biology, overview their capabilities limitations. We also speculate on possible future technological advances have potential great strides both regenerative strategies.

Language: Английский

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