Immune Cell Production of Interleukin 17 Induces Stem Cell Features of Pancreatic Intraepithelial Neoplasia Cells

Autor: Melissa Pruski, Guillermina Lozano, Sonal Gupta, Michelle Zoltan, Jennifer M. Bailey, Timothy C. Wang, Erick Riquelme, Jay K. Kolls, Paul J. Chiao, Hanwen Xu, Eduardo Vilar, Steven D. Leach, Shao Cong Sun, Anirban Maitra, William Horne, Kyle Chang, Florencia McAllister, Jianhua Ling, Susana Castro-Pando, Huamin Wang, Maria Fernanda Montiel, Yu Zhang, Ismet Sahin, Zhengyu Jiang
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Databases
Factual
Adenocarcinoma in Situ
Biology
Protein Serine-Threonine Kinases
medicine.disease_cause
Article
Aldehyde Dehydrogenase 1 Family
03 medical and health sciences
Mice
Doublecortin-Like Kinases
Pancreatic tumor
Pancreatic cancer
Pancreatitis
Chronic
medicine
Animals
Humans
Progenitor cell
Hepatology
Interleukin-17
Gastroenterology
Intracellular Signaling Peptides and Proteins
Retinal Dehydrogenase
Receptors
Interleukin
Aldehyde Dehydrogenase
medicine.disease
Flow Cytometry
Antibodies
Neutralizing
Gene Expression Regulation
Neoplastic
Pancreatic Neoplasms
030104 developmental biology
medicine.anatomical_structure
Cancer cell
Cancer research
Disease Progression
Neoplastic Stem Cells
Octamer Transcription Factors
Stem cell
Tuft cell
Pancreas
Carcinogenesis
Carcinoma
Pancreatic Ductal
Popis: BACKGROUND & AIMS: Little is known about how the immune system affects stem cell features of pancreatic cancer cells. Immune cells that produce interleukin 17 A (IL17A) in the chronically inflamed pancreas (chronic pancreatitis) contribute to PanIN initiation and progression. We investigated the effects of IL17A signaling exerts on pancreatic cancer progenitor cells and the clinical relevance of this phenomena. METHODS: We performed studies with Mist1Cre;LSLKras;Rosa26mTmG (KC(iMist);G) and Kras(G12D); Trp53(R172H); Pdx1-Cre (KPC) mice (which upon tamoxifen induction spontaneously develop pancreatic intraepithelial neoplasias, PanINs) and control littermates. Some mice were injected with neutralizing antibodies against IL17A or control antibody. Pancreata were collected, PanIN epithelial cells were isolated by flow cytometry based on lineage tracing, and gene expression profiles were compared. We collected cells from pancreatic tumors of KPC mice, incubated them with IL17 or control media, measured expression of genes regulated by IL17 signaling, injected the cancer cells into immune competent mice, and measured tumor growth. IL17A was overexpressed in pancreata of KC(iMist) mice from an adenoviral vector. Pancreata were collected from all mice and analyzed by histology and immunohistochemistry. Levels of doublecortin like kinase 1 (DCLK1) and other proteins were knocked down in KPC pancreatic cancer cells using small interfering or small hairpin RNAs; cells were analyzed by immunoblotting. We obtained 65 pancreatic tumor specimens from patients, analyzed protein levels by immunohistochemistry, and compared results with patient survival times. We also analyzed gene expression levels and patient outcome using the Cancer Genome Atlas database. RESULTS: PanIN cells from KC(iMist);G mice had a gene expression pattern associated with embryonic stem cells. Mice given injections of IL17 neutralizing antibodies, or with immune cells that did not secrete IL17, lost this expression pattern, and significantly decreased expression of DCLK1 and POU class 2 homeobox 3 (POU2F3), which regulate tuft cell development. KC(iMist) mice that overexpressed IL17 formed more PanINs, with more DCLK1-positive cells, than control mice. Pancreatic tumor cells from KPC mice and human Capan-2 cells exposed to IL17A had increased activation of NF-κB and MAPK signaling, and increased expression of DCLK1 and ALDH1A1 (a marker of embryonic stem cells), compared to cells in control media. These cells also formed tumors faster that cells not exposed to IL17 when they were injected into immunocompetent mice. KPC cells with knockdown of DCLK1 expressed lower levels of ALDH1A1 following incubation with IL17 than cells without knockdown. Expression of the IL17 receptor C (IL17RC) was higher in DCLK1-positive PanIN cells from mice compared to DCLK1-negative PanIN cells. In human pancreatic tumor tissues, high levels of DCLK1 associated with a shorter median survival time of patients (17.7 months, compared with 26.6 months of patients whose tumors had low levels of DCLK1). Tumor levels of POU2F3 and LAMC2 also associated with patient survival time. CONCLUSIONS: In studies of mouse and human pancreatic tumors and precursors, we found immune cell-derived IL17 to regulate development of tuft cells and stem cell features of pancreatic cancer cells via increased expression of DCLK1, POU2F3, ALDH1A1, and IL17RC. Strategies to disrupt this pathway might be developed to prevent pancreatic tumor growth and progression.
Databáze: OpenAIRE
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