Pancreas is a vital gland of gastrointestinal system with exocrine and endocrine secretory functions, interweaved into essential metabolic circuitries of the human body. Pancreatic ductal adenocarcinoma (PDAC) represents one of the most lethal malignancies, with a 5-year survival rate of 11%. This poor prognosis is primarily attributed to the absence of early symptoms, rapid metastatic dissemination, and the limited efficacy of current therapeutic interventions. Despite recent advancements in understanding the etiopathogenesis and treatment of PDAC, there remains a pressing need for improved individualized models, identification of novel molecular targets, and development of unbiased predictors of disease progression. Here we aim to explore the concept of precision medicine utilizing 3-dimensional, patient-specific cellular models of pancreatic tumors and discuss their potential applications in uncovering novel druggable molecular targets and predicting clinical parameters for individual patients.

Department of Biochemistry Cell and Molecular Biology 3rd Faculty of Medicine Charles University Prague

Department of Histology and Embryology Faculty of Medicine Masaryk University Brno

Departments of Internal Medicine Hematology and Oncology

International Clinical Research Center St Anne's University Hospital Brno Czech Republic

Pathology

Research Center for Applied Molecular Oncology Masaryk Memorial Cancer Institute

Surgery Clinic University Hospital Brno Faculty of Medicine Masaryk University

See more in PubMed

Cascinu S, Falconi M, Valentini V, et al; ESMO Guidelines Working Group. Pancreatic cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol . 2010;21(Suppl 5):v55–v58.

Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin . 2021;71:209–249.

Homepage—ÚZIS ČR [Internet]. Available at: https://www.uzis.cz/index-en.php . Accessed March 8, 2024.

Raffenne J, Martin FA, Nicolle R, et al. Pancreatic ductal adenocarcinoma arising in young and old patients displays similar molecular features. Cancers (Basel) . 2021;13:1234.

Stoffel EM, Brand RE, Goggins M. Pancreatic cancer: changing epidemiology and new approaches to risk assessment, early detection, and prevention. Gastroenterology . 2023;164:752–765.

Gaddam S, Abboud Y, Oh J, et al. Incidence of pancreatic cancer by age and sex in the US, 2000–2018. JAMA . 2021;326:2075–2077.

Huang BZ, Liu L, Zhang J, et al; USC Pancreas Research Team. Rising incidence and racial disparities of early-onset pancreatic cancer in the United States, 1995–2018. Gastroenterology. 2022;163:310–312.e1.

Wu W, He X, Yang L, et al. Rising trends in pancreatic cancer incidence and mortality in 2000–2014. Clin Epidemiol . 2018;10:789–797.

Siegel RL, Miller KD, Fuchs HE, et al. Cancer statistics, 2022. CA Cancer J Clin . 2022;72:7–33.

Groot VP, Rezaee N, Wu W, et al. Patterns, timing, and predictors of recurrence following pancreatectomy for pancreatic ductal adenocarcinoma. Ann Surg . 2018;267:936–945.

Poruk KE, Gay DZ, Brown K, et al. The clinical utility of CA 19-9 in pancreatic adenocarcinoma: diagnostic and prognostic updates. Curr Mol Med . 2013;13:340–351.

Kim J, Bamlet WR, Oberg AL, et al. Detection of early pancreatic ductal adenocarcinoma with thrombospondin-2 and CA19-9 blood markers. Sci Transl Med . 2017;9:eaah5583.

Ermiah E, Eddfair M, Abdulrahman O, et al. Prognostic value of serum CEA and CA19-9 levels in pancreatic ductal adenocarcinoma. Mol Clin Oncol . 2022;17:126.

Udgata S, Takenaka N, Bamlet WR, et al. THBS2/CA19-9 detecting pancreatic ductal adenocarcinoma at diagnosis underperforms in prediagnostic detection: implications for biomarker advancement. Cancer Prev Res (Phila) . 2021;14:223–232.

Seimiya T, Otsuka M, Fujishiro M. Roles of circular RNAs in the pathogenesis and treatment of pancreatic cancer. Front Cell Dev Biol . 2022;10:1023332.

Eid M, Karousi P, Kunovský L, et al. The role of circulating microRNAs in patients with early-stage pancreatic adenocarcinoma. Biomedicine . 2021;9:1468.

Wolrab D, Jirásko R, Cífková E, et al. Lipidomic profiling of human serum enables detection of pancreatic cancer. Nat Commun . 2022;13:124.

Collisson EA, Sadanandam A, Olson P, et al. Subtypes of pancreatic ductal adenocarcinoma and their differing responses to therapy. Nat Med . 2011;17:500–503.

Bailey P, Chang DK, Nones K, et al. Genomic analyses identify molecular subtypes of pancreatic cancer. Nature . 2016;531:47–52.

Sivapalan L, Kocher HM, Ross-Adams H, et al. The molecular landscape of pancreatic ductal adenocarcinoma. Pancreatology . 2022;22:925–936.

Benzel J, Fendrich V. Familial pancreatic cancer. Oncol Res Treat . 2018;41:611–618.

Roberts NJ, Norris AL, Petersen GM, et al. Whole genome sequencing defines the genetic heterogeneity of familial pancreatic cancer. Cancer Discov . 2016;6:166–175.

Aguirre AJ, Nowak JA, Camarda ND, et al. Real-time genomic characterization of advanced pancreatic cancer to enable precision medicine. Cancer Discov . 2018;8:1096–1111.

Lee JJ, Bernard V, Semaan A, et al. Elucidation of tumor-stromal heterogeneity and the ligand-receptor interactome by single-cell transcriptomics in real-world pancreatic cancer biopsies. Clin Cancer Res . 2021;27:5912–5921.

Oh K, Yoo YJ, Torre-Healy LA, et al. Coordinated single-cell tumor microenvironment dynamics reinforce pancreatic cancer subtype. Nat Commun . 2023;14:5226.

Li MM, Datto M, Duncavage EJ, et al. Standards and guidelines for the interpretation and reporting of sequence variants in cancer: a joint consensus recommendation of the Association for Molecular Pathology, American Society of Clinical Oncology, and College of American Pathologists. J Mol Diagn . 2017;19:4–23.

Almoguera C, Shibata D, Forrester K, et al. Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes. Cell . 1988;53:549–554.

Matsuda Y, Ishiwata T, Izumiyama-Shimomura N, et al. Gradual telomere shortening and increasing chromosomal instability among PanIN grades and normal ductal epithelia with and without cancer in the pancreas. PLoS One . 2015;10:e0117575.

Campa D, Matarazzi M, Greenhalf W, et al. Genetic determinants of telomere length and risk of pancreatic cancer: a PANDoRA study. Int J Cancer . 2019;144:1275–1283.

Hruban RH, Takaori K, Klimstra DS, et al. An illustrated consensus on the classification of pancreatic intraepithelial neoplasia and intraductal papillary mucinous neoplasms. Am J Surg Pathol . 2004;28:977–987.

Oldfield LE, Connor AA, Gallinger S. Molecular events in the natural history of pancreatic cancer. Trends Cancer . 2017;3:336–346.

Navas C, Hernández-Porras I, Schuhmacher AJ, et al. EGF receptor signaling is essential for k-ras oncogene-driven pancreatic ductal adenocarcinoma. Cancer Cell . 2012;22:318–330.

Ardito CM, Grüner BM, Takeuchi KK, et al. EGF receptor is required for KRAS-induced pancreatic tumorigenesis. Cancer Cell . 2012;22:304–317.

Rozenblum E, Schutte M, Goggins M, et al. Tumor-suppressive pathways in pancreatic carcinoma. Cancer Res . 1997;57:1731–1734.

Hezel AF, Kimmelman AC, Stanger BZ, et al. Genetics and biology of pancreatic ductal adenocarcinoma. Genes Dev . 2006;20:1218–1249.

Jones S, Zhang X, Parsons DW, et al. Core signaling pathways in human pancreatic cancers revealed by global genomic analyses. Science . 2008;321:1801–1806.

Iacobuzio-Donahue CA, Velculescu VE, Wolfgang CL, et al. Genetic basis of pancreas cancer development and progression: insights from whole-exome and whole-genome sequencing. Clin Cancer Res . 2012;18:4257–4265.

Waddell N, Pajic M, Patch AM, et al. Whole genomes redefine the mutational landscape of pancreatic cancer. Nature . 2015;518:495–501.

Cancer Genome Atlas Research Network. Electronic address: andrew_aguirre@dfci.harvard.edu , Cancer Genome Atlas Research Network. Integrated genomic characterization of pancreatic ductal adenocarcinoma. Cancer Cell . 2017;32:185–203.e13.

Halbrook CJ, Lyssiotis CA, Pasca di Magliano M, et al. Pancreatic cancer: advances and challenges. Cell . 2023;186:1729–1754.

Yachida S, Jones S, Bozic I, et al. Distant metastasis occurs late during the genetic evolution of pancreatic cancer. Nature . 2010;467:1114–1117.

Tian C, Clauser KR, Öhlund D, et al. Proteomic analyses of ECM during pancreatic ductal adenocarcinoma progression reveal different contributions by tumor and stromal cells. Proc Natl Acad Sci U S A . 2019;116:19609–19618.

Andrianifahanana M, Moniaux N, Schmied BM, et al. Mucin (MUC) gene expression in human pancreatic adenocarcinoma and chronic pancreatitis: a potential role of MUC4 as a tumor marker of diagnostic significance. Clin Cancer Res . 2001;7:4033–4040.

Pai P, Rachagani S, Lakshmanan I, et al. The canonical Wnt pathway regulates the metastasis-promoting mucin MUC4 in pancreatic ductal adenocarcinoma. Mol Oncol . 2016;10:224–239.

Motoda N, Matsuda Y, Onda M, et al. Overexpression of fibroblast growth factor receptor 4 in high-grade pancreatic intraepithelial neoplasia and pancreatic ductal adenocarcinoma. Int J Oncol . 2011;38:133–143.

Halbrook CJ, Lyssiotis CA. Employing metabolism to improve the diagnosis and treatment of pancreatic cancer. Cancer Cell . 2017;31:5–19.

Zuzčák M, Trnka J. Cellular metabolism in pancreatic cancer as a tool for prognosis and treatment (review). Int J Oncol . 2022;61:93.

Storz P. Acinar cell plasticity and development of pancreatic ductal adenocarcinoma. Nat Rev Gastroenterol Hepatol . 2017;14:296–304.

Reichert M, Bakir B, Moreira L, et al. Regulation of epithelial plasticity determines metastatic organotropism in pancreatic cancer. Dev Cell . 2018;45:696–711.e8.

Najafi M, Mortezaee K, Ahadi R. Cancer stem cell (a)symmetry & plasticity: tumorigenesis and therapy relevance. Life Sci . 2019;231:116520.

Boyd LNC, Andini KD, Peters GJ, et al. Heterogeneity and plasticity of cancer-associated fibroblasts in the pancreatic tumor microenvironment. Semin Cancer Biol . 2022;82:184–196.

Chen X, Song E. Turning foes to friends: targeting cancer-associated fibroblasts. Nat Rev Drug Discov . 2019;18:99–115.

Lee AYL, Dubois CL, Sarai K, et al. Cell of origin affects tumour development and phenotype in pancreatic ductal adenocarcinoma. Gut . 2019;68:487–498.

Flowers BM, Xu H, Mulligan AS, et al. Cell of origin influences pancreatic cancer subtype. Cancer Discov . 2021;11:660–677.

Mahadevan D, Von Hoff DD. Tumor-stroma interactions in pancreatic ductal adenocarcinoma. Mol Cancer Ther . 2007;6:1186–1197.

Vonlaufen A, Joshi S, Qu C, et al. Pancreatic stellate cells: partners in crime with pancreatic cancer cells. Cancer Res . 2008;68:2085–2093.

Zhang T, Ren Y, Yang P, et al. Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma. Cell Death Dis . 2022;13:897.

Nielsen MFB, Mortensen MB, Detlefsen S. Key players in pancreatic cancer-stroma interaction: cancer-associated fibroblasts, endothelial and inflammatory cells. World J Gastroenterol . 2016;22:2678–2700.

Pothula SP, Pirola RC, Wilson JS, et al. Pancreatic stellate cells: aiding and abetting pancreatic cancer progression. Pancreatology . 2020;20:409–418.

Nielsen MFB, Mortensen MB, Sørensen MD, et al. Spatial and phenotypic characterization of pancreatic cancer–associated fibroblasts after neoadjuvant treatment. Histol Histopathol . 2020;35:811–825.

Nielsen MFB, Mortensen MB, Detlefsen S. Typing of pancreatic cancer-associated fibroblasts identifies different subpopulations. World J Gastroenterol . 2018;24:4663–4678.

Biffi G, Oni TE, Spielman B, et al. IL1-induced JAK/STAT signaling is antagonized by TGFβ to shape CAF heterogeneity in pancreatic ductal adenocarcinoma. Cancer Discov . 2019;9:282–301.

Öhlund D, Handly-Santana A, Biffi G, et al. Distinct populations of inflammatory fibroblasts and myofibroblasts in pancreatic cancer. J Exp Med . 2017;214:579–596.

Goumas FA, Holmer R, Egberts JH, et al. Inhibition of IL-6 signaling significantly reduces primary tumor growth and recurrencies in orthotopic xenograft models of pancreatic cancer. Int J Cancer . 2015;137:1035–1046.

Mortezaee K. Enriched cancer stem cells, dense stroma, and cold immunity: interrelated events in pancreatic cancer. J Biochem Mol Toxicol . 2021;35:e22708.

Bhattacharjee S, Hamberger F, Ravichandra A, et al. Tumor restriction by type I collagen opposes tumor-promoting effects of cancer-associated fibroblasts. J Clin Invest . 2021;131:e146987.

Rhim AD, Oberstein PE, Thomas DH, et al. Stromal elements act to restrain, rather than support, pancreatic ductal adenocarcinoma. Cancer Cell . 2014;25:735–747.

Chen Y, Kim J, Yang S, et al. Type I collagen deletion in αSMA+ myofibroblasts augments immune suppression and accelerates progression of pancreatic cancer. Cancer Cell . 2021;39:548–565.e6.

Elyada E, Bolisetty M, Laise P, et al. Cross-species single-cell analysis of pancreatic ductal adenocarcinoma reveals antigen-presenting cancer-associated fibroblasts. Cancer Discov . 2019;9:1102–1123.

Begum A, McMillan RH, Chang YT, et al. Direct interactions with cancer-associated fibroblasts lead to enhanced pancreatic cancer stem cell function. Pancreas . 2019;48:329–334.

Ligorio M, Sil S, Malagon-Lopez J, et al. Stromal microenvironment shapes the intratumoral architecture of pancreatic cancer. Cell . 2019;178:160–175.e27.

Waghray M, Yalamanchili M, Dziubinski M, et al. GM-CSF mediates mesenchymal-epithelial cross-talk in pancreatic cancer. Cancer Discov . 2016;6:886–899.

Hermann PC, Huber SL, Herrler T, et al. Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell . 2007;1:313–323.

Maeda S, Shinchi H, Kurahara H, et al. CD133 expression is correlated with lymph node metastasis and vascular endothelial growth factor-C expression in pancreatic cancer. Br J Cancer . 2008;98:1389–1397.

Wu J, Liu Y, Fu Q, et al. Characterization of tumor-associated endothelial cells and the development of a prognostic model in pancreatic ductal adenocarcinoma. Biochim Biophys Acta Gen Subj . 1868;2024:130545.

Danilova L, Ho WJ, Zhu Q, et al. Programmed cell death ligand-1 (PD-L1) and CD8 expression profiling identify an immunologic subtype of pancreatic ductal adenocarcinomas with favorable survival. Cancer Immunol Res . 2019;7:886–895.

Helm O, Held-Feindt J, Grage-Griebenow E, et al. Tumor-associated macrophages exhibit pro- and anti-inflammatory properties by which they impact on pancreatic tumorigenesis. Int J Cancer . 2014;135:843–861.

Yang Y, Guo Z, Chen W, et al. M2 macrophage-derived exosomes promote angiogenesis and growth of pancreatic ductal adenocarcinoma by targeting E2F2. Mol Ther . 2021;29:1226–1238.

Väyrynen SA, Zhang J, Yuan C, et al. Composition, spatial characteristics, and prognostic significance of myeloid cell infiltration in pancreatic cancer. Clin Cancer Res . 2021;27:1069–1081.

Kemp SB, Pasca di Magliano M, Crawford HC. Myeloid cell mediated immune suppression in pancreatic cancer. Cell Mol Gastroenterol Hepatol . 2021;12:1531–1542.

Zhang Y, Lazarus J, Steele NG, et al. Regulatory T-cell depletion alters the tumor microenvironment and accelerates pancreatic carcinogenesis. Cancer Discov . 2020;10:422–439.

Ajina R, Weiner LM. T-cell immunity in pancreatic cancer. Pancreas . 2020;49:1014–1023.

Minici C, Rigamonti E, Lanzillotta M, et al. B lymphocytes contribute to stromal reaction in pancreatic ductal adenocarcinoma. Onco Targets Ther . 2020;9:1794359.

Raber P, Ochoa AC, Rodríguez PC. Metabolism of l -arginine by myeloid-derived suppressor cells in cancer: mechanisms of T cell suppression and therapeutic perspectives. Immunol Invest . 2012;41(6–7):614–634.

Pylayeva-Gupta Y, Das S, Handler JS, et al. IL35-producing B cells promote the development of pancreatic neoplasia. Cancer Discov . 2016;6:247–255.

Gunderson AJ, Kaneda MM, Tsujikawa T, et al. Bruton tyrosine kinase–dependent immune cell cross-talk drives pancreas cancer. Cancer Discov . 2016;6:270–285.

Mirlekar B, Wang Y, Li S, et al. Balance between immunoregulatory B cells and plasma cells drives pancreatic tumor immunity. Cell Rep Med . 2022;3:100744.

Mirlekar B, Michaud D, Lee SJ, et al. B cell-derived IL35 drives STAT3-dependent CD8 + T-cell exclusion in pancreatic cancer. Cancer Immunol Res . 2020;8:292–308.

Zhu H, Xu J, Wang W, et al. Intratumoral CD38 + CD19 + B cells associate with poor clinical outcomes and immunosuppression in patients with pancreatic ductal adenocarcinoma. EBioMedicine . 2024;103:105098.

Biondani G, Zeeberg K, Greco MR, et al. Extracellular matrix composition modulates PDAC parenchymal and stem cell plasticity and behavior through the secretome. FEBS J . 2018;285:2104–2124.

Begum A, Ewachiw T, Jung C, et al. The extracellular matrix and focal adhesion kinase signaling regulate cancer stem cell function in pancreatic ductal adenocarcinoma. PLoS One . 2017;12:e0180181.

Kim PK, Halbrook CJ, Kerk SA, et al. Hyaluronic acid fuels pancreatic cancer cell growth. Elife . 2021;10:e62645.

Janmey PA, Fletcher DA, Reinhart-King CA. Stiffness sensing by cells. Physiol Rev . 2020;100:695–724.

Hosein AN, Brekken RA, Maitra A. Pancreatic cancer stroma: an update on therapeutic targeting strategies. Nat Rev Gastroenterol Hepatol . 2020;17:487–505.

Carvalho TMA, Di Molfetta D, Greco MR, et al. Tumor microenvironment features and chemoresistance in pancreatic ductal adenocarcinoma: insights into targeting physicochemical barriers and metabolism as therapeutic approaches. Cancers (Basel) . 2021;13:6135.

Wang WQ, Liu L, Xu HX, et al. Infiltrating immune cells and gene mutations in pancreatic ductal adenocarcinoma. Br J Surg . 2016;103:1189–1199.

Kong B, Cheng T, Wu W, et al. Hypoxia-induced endoplasmic reticulum stress characterizes a necrotic phenotype of pancreatic cancer. Oncotarget . 2015;6:32154–32160.

Yamasaki A, Yanai K, Onishi H. Hypoxia and pancreatic ductal adenocarcinoma. Cancer Lett . 2020;484:9–15.

Tao J, Yang G, Zhou W, et al. Targeting hypoxic tumor microenvironment in pancreatic cancer. J Hematol Oncol . 2021;14:14.

Wishart G, Gupta P, Nisbet A, et al. On the evaluation of a novel hypoxic 3D pancreatic cancer model as a tool for radiotherapy treatment screening. Cancers (Basel) . 2021;13:6080.

Cao J, Li J, Sun L, et al. Hypoxia-driven paracrine osteopontin/integrin αvβ3 signaling promotes pancreatic cancer cell epithelial-mesenchymal transition and cancer stem cell–like properties by modulating forkhead box protein M1. Mol Oncol . 2019;13:228–245.

Schwörer S, Cimino FV, Ros M, et al. Hypoxia potentiates the inflammatory fibroblast phenotype promoted by pancreatic cancer cell–derived cytokines. Cancer Res . 2023;83:1596–1610.

Nakamura H, Takada K. Reactive oxygen species in cancer: current findings and future directions. Cancer Sci . 2021;112:3945–3952.

Liu Q, Ge W, Martínez-Jarquín S, et al. Mass spectrometry reveals high levels of hydrogen peroxide in pancreatic cancer cells. Angew Chem Int Ed Engl . 2023;62:e202213703.

Chen Z, Ho IL, Soeung M, et al. Ether phospholipids are required for mitochondrial reactive oxygen species homeostasis. Nat Commun . 2023;14:2194.

Ohara Y, Valenzuela P, Hussain SP. The interactive role of inflammatory mediators and metabolic reprogramming in pancreatic cancer. Trends Cancer . 2022;8:556–569.

Kremer DM, Nelson BS, Lin L, et al. GOT1 inhibition promotes pancreatic cancer cell death by ferroptosis. Nat Commun . 2021;12:4860.

Lopez-Blazquez C, Lacalle-Gonzalez C, Sanz-Criado L, et al. Iron-dependent cell death: a new treatment approach against pancreatic ductal adenocarcinoma. Int J Mol Sci . 2023;24:14979.

Huang X, He C, Hua X, et al. Oxidative stress induces monocyte-to-myofibroblast transdifferentiation through p38 in pancreatic ductal adenocarcinoma. Clin Transl Med . 2020;10:e41.

Lee KE, Spata M, Bayne LJ, et al. Hif1a deletion reveals pro-neoplastic function of B cells in pancreatic neoplasia. Cancer Discov . 2016;6:256–269.

Longnecker DS. Anatomy and Histology of the Pancreas. Pancreapedia: The Exocrine Pancreas Knowledge Base [Internet]. 2021. Available at: https://www.pancreapedia.org/reviews/anatomy-and-histology-of-pancreas . Accessed February 29, 2024.

Pan FC, Brissova M. Pancreas development in humans. Curr Opin Endocrinol Diabetes Obes . 2014;21:77–82.

Jennings RE, Berry AA, Kirkwood-Wilson R, et al. Development of the human pancreas from foregut to endocrine commitment. Diabetes . 2013;62:3514–3522.

Tiso N, Moro E, Argenton F. Zebrafish pancreas development. Mol Cell Endocrinol . 2009;312(1–2):24–30.

Jørgensen MC, Ahnfelt-Rønne J, Hald J, et al. An illustrated review of early pancreas development in the mouse. Endocr Rev . 2007;28:685–705.

McCracken KW, Wells JM. Molecular pathways controlling pancreas induction. Semin Cell Dev Biol . 2012;23:656–662.

Bastidas-Ponce A, Scheibner K, Lickert H, et al. Cellular and molecular mechanisms coordinating pancreas development. Development . 2017;144:2873–2888.

Hebrok M. Hedgehog signaling in pancreas development. Mech Dev . 2003;120:45–57.

Mfopou JK, De Groote V, Xu X, et al. Sonic hedgehog and other soluble factors from differentiating embryoid bodies inhibit pancreas development. Stem Cells . 2007;25:1156–1165.

Hebrok M, Kim SK, Melton DA. Notochord repression of endodermal sonic hedgehog permits pancreas development. Genes Dev . 1998;12:1705–1713.

Xuan S, Sussel L. GATA4 and GATA6 regulate pancreatic endoderm identity through inhibition of hedgehog signaling. Development . 2016;143:780–786.

Shi ZD, Lee K, Yang D, et al. Genome editing in hPSCs reveals GATA6 haploinsufficiency and a genetic interaction with GATA4 in human pancreatic development. Cell Stem Cell . 2017;20:675–688.e6.

Murtaugh LC, Melton DA. Genes, signals, and lineages in pancreas development. Annu Rev Cell Dev Biol . 2003;19:71–89.

Apelqvist A, Li H, Sommer L, et al. Notch signalling controls pancreatic cell differentiation. Nature . 1999;400:877–881.

Baumgartner BK, Cash G, Hansen H, et al. Distinct requirements for beta-catenin in pancreatic epithelial growth and patterning. Dev Biol . 2014;391:89–98.

Scheibner K, Bakhti M, Bastidas-Ponce A, et al. Wnt signaling: implications in endoderm development and pancreas organogenesis. Curr Opin Cell Biol . 2019;61:48–55.

Murtaugh LC, Law AC, Dor Y, et al. Beta-catenin is essential for pancreatic acinar but not islet development. Development . 2005;132:4663–4674.

Miralles F, Czernichow P, Scharfmann R. Follistatin regulates the relative proportions of endocrine versus exocrine tissue during pancreatic development. Development . 1998;125:1017–1024.

Ma Z, Zhang X, Zhong W, et al. Deciphering early human pancreas development at the single-cell level. Nat Commun . 2023;14:5354.

Tosti L, Hang Y, Debnath O, et al. Single-nucleus and in situ RNA-sequencing reveal cell topographies in the human pancreas. Gastroenterology . 2021;160:1330–1344.e11.

Dettmer R, Cirksena K, Münchhoff J, et al. FGF2 inhibits early pancreatic lineage specification during differentiation of human embryonic stem cells. Cells . 2020;9:1927.

Sui L, Geens M, Sermon K, et al. Role of BMP signaling in pancreatic progenitor differentiation from human embryonic stem cells. Stem Cell Rev Rep . 2013;9:569–577.

Takeuchi K, Tabe S, Takahashi K, et al. Incorporation of human iPSC-derived stromal cells creates a pancreatic cancer organoid with heterogeneous cancer-associated fibroblasts. Cell Rep . 2023;42:113420.

Yu Y, Yang G, Huang H, et al. Preclinical models of pancreatic ductal adenocarcinoma: challenges and opportunities in the era of precision medicine. J Exp Clin Cancer Res . 2021;40:8.

Kim J, Hoffman JP, Alpaugh RK, et al. An iPSC line from human pancreatic ductal adenocarcinoma undergoes early to invasive stages of pancreatic cancer progression. Cell Rep . 2013;3:2088–2099.

Kim J, Zaret KS. Generation of induced pluripotent stem cell–like lines from human pancreatic ductal adenocarcinoma. Methods Mol Biol . 1882;2019:33–53.

Simsek S, Zhou T, Robinson CL, et al. Modeling cystic fibrosis using pluripotent stem cell–derived human pancreatic ductal epithelial cells. Stem Cells Transl Med . 2016;5:572–579.

Petersen MBK, Gonçalves CAC, Kim YH, et al. Recapitulating and deciphering human pancreas development from human pluripotent stem cells in a dish. Curr Top Dev Biol . 2018;129:143–190.

Seino T, Kawasaki S, Shimokawa M, et al. Human pancreatic tumor organoids reveal loss of stem cell niche factor dependence during disease progression. Cell Stem Cell . 2018;22:454–467.e6.

Pagliuca FW, Millman JR, Gürtler M, et al. Generation of functional human pancreatic β cells in vitro. Cell . 2014;159:428–439.

Maloy MH, Ferrer MA, Parashurama N. In vivo differentiation of stem cell-derived human pancreatic progenitors to treat type 1 diabetes. Stem Cell Rev Rep . 2020;16:1139–1155.

Thakur G, Lee HJ, Jeon RH, et al. Small molecule-induced pancreatic β-like cell development: mechanistic approaches and available strategies. Int J Mol Sci . 2020;21:2388.

Huang L, Holtzinger A, Jagan I, et al. Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell– and patient-derived tumor organoids. Nat Med . 2015;21:1364–1371.

Kokkinos J, Sharbeen G, Haghighi KS, et al. Ex vivo culture of intact human patient derived pancreatic tumour tissue. Sci Rep . 2021;11:1944.

Guerra C, Barbacid M. Genetically engineered mouse models of pancreatic adenocarcinoma. Mol Oncol . 2013;7:232–247.

Kim MP, Evans DB, Wang H, et al. Generation of orthotopic and heterotopic human pancreatic cancer xenografts in immunodeficient mice. Nat Protoc . 2009;4:1670–1680.

Mallya K, Gautam SK, Aithal A, et al. Modeling pancreatic cancer in mice for experimental therapeutics. Biochim Biophys Acta Rev Cancer . 1876;2021:188554.

Sant S, Johnston PA. The production of 3D tumor spheroids for cancer drug discovery. Drug Discov Today Technol . 2017;23:27–36.

Longati P, Jia X, Eimer J, et al. 3D pancreatic carcinoma spheroids induce a matrix-rich, chemoresistant phenotype offering a better model for drug testing. BMC Cancer . 2013;13:95.

Mehta G, Hsiao AY, Ingram M, et al. Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. J Control Release . 2012;164:192–204.

Braun LM, Lagies S, Klar RFU, et al. Metabolic profiling of early and late recurrent pancreatic ductal adenocarcinoma using patient-derived organoid cultures. Cancers (Basel) . 2020;12:1440.

Sharick JT, Walsh CM, Sprackling CM, et al. Metabolic heterogeneity in patient tumor-derived organoids by primary site and drug treatment. Front Oncol . 2020;10:553.

Huang B, Wei X, Chen K, et al. Bioprinting of hydrogel beads to engineer pancreatic tumor-stroma microtissues for drug screening. Int J Bioprint . 2023;9:676.

Geyer M, Gaul LM, D'Agosto SL, et al. The tumor stroma influences immune cell distribution and recruitment in a PDAC-on-a-chip model. Front Immunol . 2023;14:1155085.

Haque MR, Wessel CR, Leary DD, et al. Patient-derived pancreatic cancer-on-a-chip recapitulates the tumor microenvironment. Microsyst Nanoeng . 2022;8:36.

Geyer M, Schreyer D, Gaul LM, et al. A microfluidic-based PDAC organoid system reveals the impact of hypoxia in response to treatment. Cell Death Dis . 2023;9:20.

Deer EL, González-Hernández J, Coursen JD, et al. Phenotype and genotype of pancreatic cancer cell lines. Pancreas . 2010;39:425–435.

Monberg ME, Geiger H, Lee JJ, et al. Occult polyclonality of preclinical pancreatic cancer models drives in vitro evolution. Nat Commun . 2022;13:3652.

Daniel SK, Sullivan KM, Labadie KP, et al. Hypoxia as a barrier to immunotherapy in pancreatic adenocarcinoma. Clin Transl Med . 2019;8:10.

Shoval H, Karsch-Bluman A, Brill-Karniely Y, et al. Tumor cells and their crosstalk with endothelial cells in 3D spheroids. Sci Rep . 2017;7:10428.

Timmins NE, Nielsen LK. Generation of multicellular tumor spheroids by the hanging-drop method. Methods Mol Med . 2007;140:141–151.

Ware MJ, Colbert K, Keshishian V, et al. Generation of homogenous three-dimensional pancreatic cancer cell spheroids using an improved hanging drop technique. Tissue Eng Part C Methods . 2016;22:312–321.

Ware MJ, Keshishian V, Law JJ, et al. Generation of an in vitro 3D PDAC stroma rich spheroid model. Biomaterials . 2016;108:129–142.

Giustarini G, Teng G, Pavesi A, et al. Characterization of 3D heterocellular spheroids of pancreatic ductal adenocarcinoma for the study of cell interactions in the tumor immune microenvironment. Front Oncol . 2023;13:1156769.

Tomas Bort E, Joseph MD, Wang Q, et al. Purinergic GPCR-integrin interactions drive pancreatic cancer cell invasion. Elife . 2023;12:e86971.

Saad MA, Zhung W, Stanley ME, et al. Photoimmunotherapy retains its anti-tumor efficacy with increasing stromal content in heterotypic pancreatic cancer spheroids. Mol Pharm . 2022;19:2549–2563.

Sarhan D, Eisinger S, He F, et al. Targeting myeloid suppressive cells revives cytotoxic anti-tumor responses in pancreatic cancer. iScience . 2022;25:105317.

Czaplinska D, Ialchina R, Andersen HB, et al. Crosstalk between tumor acidosis, p53 and extracellular matrix regulates pancreatic cancer aggressiveness. Int J Cancer . 2023;152:1210–1225.

Daunke T, Beckinger S, Rahn S, et al. Expression and role of the immune checkpoint regulator PD-L1 in the tumor-stroma interplay of pancreatic ductal adenocarcinoma. Front Immunol . 2023;14:1157397.

McKenna MK, Ozcan A, Brenner D, et al. Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma. J Immunother Cancer . 2023;11:e005891.

Tidwell TR, Røsland GV, Tronstad KJ, et al. Metabolic flux analysis of 3D spheroids reveals significant differences in glucose metabolism from matched 2D cultures of colorectal cancer and pancreatic ductal adenocarcinoma cell lines. Cancer Metab . 2022;10:9.

Vakhshiteh F, Bagheri Z, Soleimani M, et al. Heterotypic tumor spheroids: a platform for nanomedicine evaluation. J Nanobiotechnology . 2023;21:249.

Ermis M, Falcone N, Roberto de Barros N, et al. Tunable hybrid hydrogels with multicellular spheroids for modeling desmoplastic pancreatic cancer. Bioact Mater . 2023;25:360–373.

Shichi Y, Sasaki N, Michishita M, et al. Enhanced morphological and functional differences of pancreatic cancer with epithelial or mesenchymal characteristics in 3D culture. Sci Rep . 2019;9:10871.

Minami F, Sasaki N, Shichi Y, et al. Morphofunctional analysis of human pancreatic cancer cell lines in 2- and 3-dimensional cultures. Sci Rep . 2021;11:6775.

Beer M, Kuppalu N, Stefanini M, et al. A novel microfluidic 3D platform for culturing pancreatic ductal adenocarcinoma cells: comparison with in vitro cultures and in vivo xenografts. Sci Rep . 2017;7:1325.

Damghani T, Moosavi F, Khoshneviszadeh M, et al. Imidazopyridine hydrazone derivatives exert antiproliferative effect on lung and pancreatic cancer cells and potentially inhibit receptor tyrosine kinases including c-Met. Sci Rep . 2021;11:3644.

Mortazavi M, Raufi E, Damghani T, et al. Discovery of anticancer agents with c-Met inhibitory potential by virtual and experimental screening of a chemical library. Eur J Pharmacol . 2023;938:175395.

Bhattacharyya S, Ghosh H, Covarrubias-Zambrano O, et al. Anticancer activity of novel difluorinated curcumin analog and its inclusion complex with 2-hydroxypropyl-β-cyclodextrin against pancreatic cancer. Int J Mol Sci . 2023;24:6336.

Drifka CR, Eliceiri KW, Weber SM, et al. A bioengineered heterotypic stroma-cancer microenvironment model to study pancreatic ductal adenocarcinoma. Lab Chip . 2013;13:3965–3975.

Lee JH, Kim SK, Khawar IA, et al. Microfluidic co-culture of pancreatic tumor spheroids with stellate cells as a novel 3D model for investigation of stroma-mediated cell motility and drug resistance. J Exp Clin Cancer Res . 2018;37:4.

Brumskill S, Barrera LN, Calcraft P, et al. Inclusion of cancer-associated fibroblasts in drug screening assays to evaluate pancreatic cancer resistance to therapeutic drugs. J Physiol Biochem . 2023;79:223–234.

Hingorani SR, Zheng L, Bullock AJ, et al. HALO 202: randomized phase II study of PEGPH20 plus nab-paclitaxel/gemcitabine versus nab-paclitaxel/gemcitabine in patients with untreated, metastatic pancreatic ductal adenocarcinoma. J Clin Oncol . 2018;36:359–366.

Ramanathan RK, McDonough SL, Philip PA, et al. Phase IB/II randomized study of FOLFIRINOX plus pegylated recombinant human hyaluronidase versus FOLFIRINOX alone in patients with metastatic pancreatic adenocarcinoma: SWOG S1313. J Clin Oncol . 2019;37:1062–1069.

Kuninty PR, Bansal R, De Geus SWL, et al. ITGA5 inhibition in pancreatic stellate cells attenuates desmoplasia and potentiates efficacy of chemotherapy in pancreatic cancer. Sci Adv . 2019;5:eaax2770.

Kuninty PR, Bojmar L, Tjomsland V, et al. MicroRNA-199a and -214 as potential therapeutic targets in pancreatic stellate cells in pancreatic tumor. Oncotarget . 2016;7:16396–16408.

Bots STF, Harryvan TJ, Groeneveldt C, et al. Preclinical evaluation of the gorilla-derived HAdV-B AdV-lumc007 oncolytic adenovirus ‘GoraVir’ for the treatment of pancreatic ductal adenocarcinoma. Mol Oncol . 2024;18:1245–1258.

Lazzari G, Nicolas V, Matsusaki M, et al. Multicellular spheroid based on a triple co-culture: a novel 3D model to mimic pancreatic tumor complexity. Acta Biomater . 2018;78:296–307.

Kpeglo D, Hughes MDG, Dougan L, et al. Modeling the mechanical stiffness of pancreatic ductal adenocarcinoma. Matrix Biol Plus . 2022;14:100109.

Sato T, Clevers H. Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications. Science . 2013;340:1190–1194.

Boj SF, Hwang CI, Baker LA, et al. Organoid models of human and mouse ductal pancreatic cancer. Cell . 2015;160(1–2):324–338.

Driehuis E, Gracanin A, Vries RGJ, et al. Establishment of pancreatic organoids from Normal tissue and tumors. STAR Protoc . 2020;1:100192.

Shi X, Li Y, Yuan Q, et al. Integrated profiling of human pancreatic cancer organoids reveals chromatin accessibility features associated with drug sensitivity. Nat Commun . 2022;13:2169.

VanderLaan PA. Fine-needle aspiration and core needle biopsy: an update on 2 common minimally invasive tissue sampling modalities. Cancer Cytopathol . 2016;124:862–870.

Sagami R, Nakahodo J, Minami R, et al. True diagnostic ability of EUS-guided fine-needle aspiration/biopsy sampling for small pancreatic lesions ≤10 mm and salvage diagnosis by pancreatic juice cytology: a multicenter study. Gastrointest Endosc . 2024;99:73–80.

Heckler M, Hackert T. Surgery for locally advanced pancreatic ductal adenocarcinoma—is it only about the vessels? J Gastrointest Oncol . 2021;12:2503–2511.

Driehuis E, van Hoeck A, Moore K, et al. Pancreatic cancer organoids recapitulate disease and allow personalized drug screening. Proc Natl Acad Sci U S A . 2019;116:26580–26590.

Baker LA, Tiriac H, Tuveson DA. Generation and culture of human pancreatic ductal adenocarcinoma organoids from resected tumor specimens. Methods Mol Biol . 2019;1882:97–115.

Ehlen L, Arndt J, Treue D, et al. Novel methods for in vitro modeling of pancreatic cancer reveal important aspects for successful primary cell culture. BMC Cancer . 2020;20:417.

Choi JI, Jang SI, Hong J, et al. Cancer-initiating cells in human pancreatic cancer organoids are maintained by interactions with endothelial cells. Cancer Lett . 2021;498:42–53.

Romero-Calvo I, Weber CR, Ray M, et al. Human organoids share structural and genetic features with primary pancreatic adenocarcinoma tumors. Mol Cancer Res . 2019;17:70–83.

Baker LA, Tiriac H, Clevers H, et al. Modeling pancreatic cancer with organoids. Trends Cancer . 2016;2:176–190.

Walsh AJ, Castellanos JA, Nagathihalli NS, et al. Optical imaging of drug-induced metabolism changes in murine and human pancreatic cancer organoids reveals heterogeneous drug response. Pancreas . 2016;45:863–869.

Tsai S, McOlash L, Palen K, et al. Development of primary human pancreatic cancer organoids, matched stromal and immune cells and 3D tumor microenvironment models. BMC Cancer . 2018;18:335.

Tiriac H, Belleau P, Engle DD, et al. Organoid profiling identifies common responders to chemotherapy in pancreatic cancer. Cancer Discov . 2018;8:1112–1129.

Zeeberg K, Cardone RA, Greco MR, et al. Assessment of different 3D culture systems to study tumor phenotype and chemosensitivity in pancreatic ductal adenocarcinoma. Int J Oncol . 2016;49:243–252.

Heinrich MA, Uboldi I, Kuninty PR, et al. Microarchitectural mimicking of stroma-induced vasculature compression in pancreatic tumors using a 3D engineered model. Bioact Mater . 2023;22:18–33.

Osuna de la Peña D, Trabulo SMD, Collin E, et al. Bioengineered 3D models of human pancreatic cancer recapitulate in vivo tumour biology. Nat Commun . 2021;12:5623.

Gupta P, Pérez-Mancera PA, Kocher H, et al. A novel scaffold-based hybrid multicellular model for pancreatic ductal adenocarcinoma—toward a better mimicry of the in vivo tumor microenvironment. Front Bioeng Biotechnol . 2020;8:290.

Langer EM, Allen-Petersen BL, King SM, et al. Modeling tumor phenotypes in vitro with three-dimensional bioprinting. Cell Rep . 2019;26:608–623.e6.

Wei X, Wu Y, Chen K, et al. Embedded bioprinted multicellular spheroids modeling pancreatic cancer bioarchitecture towards advanced drug therapy. J Mater Chem B . 2024;12:1788–1797.

Jeong SY, Lee JH, Shin Y, et al. Co-culture of tumor spheroids and fibroblasts in a collagen matrix-incorporated microfluidic chip mimics reciprocal activation in solid tumor microenvironment. PLoS One . 2016;11:e0159013.

Kpeglo D, Haddrick M, Knowles MA, et al. Modelling and breaking down the biophysical barriers to drug delivery in pancreatic cancer. Lab Chip . 2024;24:854–868.

Kumano K, Nakahashi H, Louphrasitthiphol P, et al. Hypoxia at 3D organoid establishment selects essential subclones within heterogenous pancreatic cancer. Front Cell Dev Biol . 2024;12:1327772.

Broekgaarden M, Anbil S, Bulin AL, et al. Modulation of redox metabolism negates cancer-associated fibroblasts-induced treatment resistance in a heterotypic 3D culture platform of pancreatic cancer. Biomaterials . 2019;222:119421.