Hematopoietic progenitor kinase 1 inhibits the development and progression of pancreatic intraepithelial neoplasia

Ras plays an essential role in the development of acinar-to-ductal metaplasia (ADM) and pancreatic ductal adenocarcinoma (PDAC). However, mutant Kras is an inefficient driver for PDAC development. The mechanisms of the switching from low Ras activity to high Ras activity that are required for development and progression of pancreatic intraepithelial neoplasias (PanINs) are unclear. In this study, we found that hematopoietic progenitor kinase 1 (HPK1) was upregulated during pancreatic injury and ADM. HPK1 interacted with the SH3 domain and phosphorylated Ras GTPase-activating protein (RasGAP) and upregulated RasGAP activity. Using transgenic mouse models of HPK1 or M46, a kinase-dead mutant of HPK1, we showed that HPK1 inhibited Ras activity and its downstream signaling and regulated acinar cell plasticity. M46 promoted the development of ADM and PanINs. Expression of M46 in KrasG12D Bac mice promoted the infiltration of myeloid-derived suppressor cells and macrophages, inhibited the infiltration of T cells, and accelerated the progression of PanINs to invasive and metastatic PDAC, while HPK1 attenuated mutant Kras–driven PanIN progression. Our results showed that HPK1 plays an important role in ADM and the progression of PanINs by regulating Ras signaling. Loss of HPK1 kinase activity promotes an immunosuppressive tumor microenvironment and accelerates the progression of PanINs to PDAC.

[1]  Philip S. Yu,et al.  Single-nucleus and spatial transcriptome profiling of pancreatic cancer identifies multicellular dynamics associated with neoadjuvant treatment , 2022, Nature Genetics.

[2]  E. Cheung,et al.  Loss of the Wild-type KRAS Allele Promotes Pancreatic Cancer Progression through Functional Activation of YAP1 , 2021, Oncogene.

[3]  R. Kortum,et al.  The Role of Wild-Type RAS in Oncogenic RAS Transformation , 2021, Genes.

[4]  E. Lees,et al.  Enhanced antitumor immunity by a novel small molecule HPK1 inhibitor , 2021, Journal for ImmunoTherapy of Cancer.

[5]  Haiyan Xu,et al.  Pharmacological inhibition of hematopoietic progenitor kinase 1 positively regulates T-cell function , 2020, PloS one.

[6]  D. Dimitrov,et al.  Hematopoietic Progenitor Kinase1 (HPK1) Mediates T Cell Dysfunction and Is a Druggable Target for T Cell-Based Immunotherapies. , 2020, Cancer cell.

[7]  D. Chatterjee,et al.  Pathology of Treated Pancreatic Ductal Adenocarcinoma and Its Clinical Implications. , 2020, Archives of pathology & laboratory medicine.

[8]  A. Maitra,et al.  Hematopoietic progenitor kinase 1 down-regulates the oncogenic receptor tyrosine kinase AXL in pancreatic cancer , 2020, The Journal of Biological Chemistry.

[9]  R. Stephens,et al.  KRAS G13D sensitivity to neurofibromin-mediated GTP hydrolysis , 2019, Proceedings of the National Academy of Sciences.

[10]  R. Huganir,et al.  Tumor Suppression of Ras GTPase-Activating Protein RASA5 through Antagonizing Ras Signaling Perturbation in Carcinomas , 2019, iScience.

[11]  J. Hunt,et al.  Critical role of kinase activity of hematopoietic progenitor kinase 1 in anti-tumor immune surveillance , 2019, PloS one.

[12]  I. Mellman,et al.  The Kinase Activity of Hematopoietic Progenitor Kinase 1 Is Essential for the Regulation of T Cell Function. , 2018, Cell reports.

[13]  Liuxing Wang,et al.  HPK1 positive expression associated with longer overall survival in patients with estrogen receptor-positive invasive ductal carcinoma-not otherwise specified , 2017, Molecular medicine reports.

[14]  P. Storz.  Acinar cell plasticity and development of pancreatic ductal adenocarcinoma , 2017, Nature Reviews Gastroenterology &Hepatology.

[15]  A. Maitra,et al.  The emerging roles of F-box proteins in pancreatic tumorigenesis. , 2016, Seminars in cancer biology.

[16]  A. Maitra,et al.  Obesity, Intrapancreatic Fatty Infiltration, and Pancreatic Cancer , 2015, Clinical Cancer Research.

[17]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[18]  Haojie Huang,et al.  Oncogenic K-Ras requires activation for enhanced activity , 2014, Oncogene.

[19]  Huamin Wang,et al.  The CUL7/F-box and WD Repeat Domain Containing 8 (CUL7/Fbxw8) Ubiquitin Ligase Promotes Degradation of Hematopoietic Progenitor Kinase 1* , 2013, The Journal of Biological Chemistry.

[20]  C. Roland,et al.  A high-fat diet activates oncogenic Kras and COX2 to induce development of pancreatic ductal adenocarcinoma in mice. , 2013, Gastroenterology.

[21]  R. Pommier,et al.  Isolation and culture of mouse primary pancreatic acinar cells. , 2013, Journal of visualized experiments : JoVE.

[22]  C. Logsdon,et al.  Roles for KRAS in pancreatic tumor development and progression. , 2013, Gastroenterology.

[23]  Bronwen L. Aken,et al.  GENCODE: The reference human genome annotation for The ENCODE Project , 2012, Genome research.

[24]  Jeffrey E. Lee,et al.  Histologic grading of the extent of residual carcinoma following neoadjuvant chemoradiation in pancreatic ductal adenocarcinoma , 2012, Cancer.

[25]  Gerald C. Chu,et al.  Oncogenic Kras Maintains Pancreatic Tumors through Regulation of Anabolic Glucose Metabolism , 2012, Cell.

[26]  R. Hruban,et al.  Disruption of p16 and Activation of Kras in Pancreas Increase Ductal Adenocarcinoma Formation and Metastasis in vivo , 2011, Oncotarget.

[27]  Y. Moroi,et al.  Hematopoietic Progenitor Kinase 1, Mitogen-Activated Protein/Extracellular Signal-Related Protein Kinase Kinase Kinase 1, and phosphoMitogen-Activated Protein Kinase Kinase 4 are Overexpressed in Extramammary Paget Disease , 2011, The American Journal of dermatopathology.

[28]  M. Barbacid,et al.  Pancreatitis-induced inflammation contributes to pancreatic cancer by inhibiting oncogene-induced senescence. , 2011, Cancer cell.

[29]  N. Zaborsky,et al.  HPK1 Associates with SKAP-HOM to Negatively Regulate Rap1-Mediated B-Lymphocyte Adhesion , 2010, PloS one.

[30]  M. Hebrok,et al.  Beta-catenin blocks Kras-dependent reprogramming of acini into pancreatic cancer precursor lesions in mice. , 2010, The Journal of clinical investigation.

[31]  Gerald C. Chu,et al.  Context-dependent transformation of adult pancreatic cells by oncogenic K-Ras. , 2009, Cancer cell.

[32]  B. Ji,et al.  Ras activity in acinar cells links chronic pancreatitis and pancreatic cancer. , 2009, Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association.

[33]  Huamin Wang,et al.  Ras activity levels control the development of pancreatic diseases. , 2009, Gastroenterology.

[34]  M. Korc,et al.  Acute pancreatitis markedly accelerates pancreatic cancer progression in mice expressing oncogenic Kras. , 2009, Biochemical and biophysical research communications.

[35]  Christiane Garbay,et al.  p120-Ras GTPase activating protein (RasGAP): a multi-interacting protein in downstream signaling. , 2009, Biochimie.

[36]  Douglas B. Evans,et al.  Proteasome-mediated degradation and functions of hematopoietic progenitor kinase 1 in pancreatic cancer. , 2009, Cancer research.

[37]  G. Feldmann,et al.  Spontaneous induction of murine pancreatic intraepithelial neoplasia (mPanIN) by acinar cell targeting of oncogenic Kras in adult mice , 2008, Proceedings of the National Academy of Sciences.

[38]  B. Ji,et al.  Robust acinar cell transgene expression of CreErT via BAC recombineering , 2008, Genesis.

[39]  R. Schmid,et al.  Concomitant pancreatic activation of Kras(G12D) and Tgfa results in cystic papillary neoplasms reminiscent of human IPMN. , 2007, Cancer cell.

[40]  R. Hruban,et al.  Kras(G12D) and Smad4/Dpc4 haploinsufficiency cooperate to induce mucinous cystic neoplasms and invasive adenocarcinoma of the pancreas. , 2007, Cancer cell.

[41]  M. Barbacid,et al.  Chronic pancreatitis is essential for induction of pancreatic ductal adenocarcinoma by K-Ras oncogenes in adult mice. , 2007, Cancer cell.

[42]  A. F. Castro,et al.  Measuring Ras-family GTP levels in vivo--running hot and cold. , 2005, Methods.

[43]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[44]  R. Hruban,et al.  Trp53R172H and KrasG12D cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice. , 2005, Cancer cell.

[45]  E. Cameron,et al.  Recapitulation of elements of embryonic development in adult mouse pancreatic regeneration. , 2005, Gastroenterology.

[46]  R. DePinho,et al.  Activated Kras and Ink4a/Arf deficiency cooperate to produce metastatic pancreatic ductal adenocarcinoma. , 2003, Genes & development.

[47]  E. Petricoin,et al.  Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. , 2003, Cancer cell.

[48]  F. Brembeck,et al.  The mutant K-ras oncogene causes pancreatic periductal lymphocytic infiltration and gastric mucous neck cell hyperplasia in transgenic mice. , 2003, Cancer research.

[49]  D. Longnecker,et al.  Preinvasive pancreatic neoplasia of ductal phenotype induced by acinar cell targeting of mutant Kras in transgenic mice. , 2003, Cancer research.

[50]  G. Klöppel,et al.  Ductal lesions in patients with chronic pancreatitis show K‐ras mutations in a frequency similar to that in the normal pancreas and lack nuclear immunoreactivity for p53 , 2000, Cancer.

[51]  E. Génot,et al.  Ras regulation and function in lymphocytes. , 2000, Current opinion in immunology.

[52]  J. Van Laethem.  Ki‐Ras Oncogene Mutations in Chronic Pancreatitis: Which Discriminating Ability for Malignant Potential? , 1999, Annals of the New York Academy of Sciences.

[53]  C. Der,et al.  Increasing Complexity of the Ras Signaling Pathway* , 1998, The Journal of Biological Chemistry.

[54]  T. Pawson,et al.  SH2/SH3 Adaptor Proteins Can Link Tyrosine Kinases to a Ste20-related Protein Kinase, HPK1* , 1997, The Journal of Biological Chemistry.

[55]  T. Pawson,et al.  HPK1, a hematopoietic protein kinase activating the SAPK/JNK pathway. , 1996, The EMBO journal.

[56]  M. C. Hu,et al.  Human HPK1, a novel human hematopoietic progenitor kinase that activates the JNK/SAPK kinase cascade. , 1996, Genes & development.

[57]  D. Klimstra,et al.  K-ras mutations in pancreatic ductal proliferative lesions. , 1994, The American journal of pathology.

[58]  F. McCormick,et al.  Oncogenic and wild-type Ras play divergent roles in the regulation of mitogen-activated protein kinase signaling. , 2013, Cancer discovery.

[59]  Thomas R. Gingeras,et al.  STAR: ultrafast universal RNA-seq aligner , 2013, Bioinform..

[60]  T. Tan,et al.  Hematopoietic progenitor kinase 1 negatively regulates T cell receptor signaling and T cell–mediated immune responses , 2007, Nature Immunology.

[61]  T. Jacks,et al.  Mist1-KrasG12D knock-in mice develop mixed differentiation metastatic exocrine pancreatic carcinoma and hepatocellular carcinoma. , 2006, Cancer research.

[62]  K. Okumura,et al.  Assignment of IkappaB kinase beta (IKBKB) to human chromosome band 8p12-->p11 by in situ hybridization. , 1998, Cytogenetics and Cell Genetics.

[63]  Y. Shiratori,et al.  Analysis of K-ras gene mutation in hyperplastic duct cells of the pancreas without pancreatic disease. , 1996, Gastroenterology.

[64]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .