Notch signaling via Wnt regulates the proliferation of alternative, CCR2-independent tumor-associated macrophages in hepatocellular carcinoma.

Tumor-associated macrophages (TAMs) play pivotal roles in tumor progression and metastasis, but the contribution and regulation of different macrophage populations remain unclear. Here we show that Notch signaling plays distinct roles in regulating different TAM subsets in hepatocellular carcinoma (HCC). Myeloid-specific Notch blockade by conditional disruption of recombination signal binding protein Jϰ (RBPj cKO) significantly delayed the growth of subcutaneously inoculated Lewis lung carcinoma (LLC), but accelerated orthotopically inoculated hepatic Hepa1-6 tumors in mice. In contrast to subcutaneous LLC, RBPj cKO significantly increased the number of TAMs in hepatic Hepa1-6 tumors despite impeded differentiation of monocyte-derived TAMs (moTAMs). The dominating TAMs in orthotopic HCC manifested properties of Kupffer cells (KCs) and hence tentatively named KC-like TAMs (kclTAMs). The increased proliferation of RBPj cKO kclTAMs was maintained even in CCR2-/- mice in which moTAMs were genetically blocked. Notch signaling blockade accelerated proliferation of kclTAMs via enhanced β-catenin-dependent Wnt signaling, which also downregulated interleukin (IL)-12 and upregulated IL-10 expression by kclTAMs likely through c-Myc. Additionally, myeloid-specific RBPj cKO facilitated hepatic metastasis of colorectal cancer (CRC) but suppressed lung metastasis in mice, suggesting that the phenotype of RBPj cKO in promoting tumor growth was liver-specific. In patient-derived HCC biopsies, Notch signaling negatively correlated with Wnt activation in CD68+ macrophages, which positively correlated with advanced HCC stages. Therefore, Notch blockade impedes the differentiation of moTAMs, but upregulates Wnt/β-catenin signaling to promote the proliferation and pro-tumor cytokine production of kclTAMs, facilitating HCC progression and hepatic metastasis of CRC.

[1]  G. Storm,et al.  Inhibition of canonical WNT signaling pathway by β-catenin/CBP inhibitor ICG-001 ameliorates liver fibrosis in vivo through suppression of stromal CXCL12. , 2018, Biochimica et biophysica acta. Molecular basis of disease.

[2]  N. Carragher,et al.  Monocytes Differentiate to Immune Suppressive Precursors of Metastasis-Associated Macrophages in Mouse Models of Metastatic Breast Cancer , 2018, Front. Immunol..

[3]  Jingqin Luo,et al.  Tissue‐Resident Macrophages in Pancreatic Ductal Adenocarcinoma Originate from Embryonic Hematopoiesis and Promote Tumor Progression , 2017, Immunity.

[4]  Junwei Yang,et al.  Wnt/β-Catenin-Promoted Macrophage Alternative Activation Contributes to Kidney Fibrosis. , 2017, Journal of the American Society of Nephrology : JASN.

[5]  Darjus F. Tschaharganeh,et al.  Kupffer Cell-Derived Tnf Triggers Cholangiocellular Tumorigenesis through JNK due to Chronic Mitochondrial Dysfunction and ROS. , 2017, Cancer cell.

[6]  F. Tacke,et al.  Liver macrophages in tissue homeostasis and disease , 2017, Nature Reviews Immunology.

[7]  Alberto Mantovani,et al.  Tumour-associated macrophages as treatment targets in oncology , 2017, Nature Reviews Clinical Oncology.

[8]  Jane E. Dalton,et al.  Bone marrow-derived and resident liver macrophages display unique transcriptomic signatures but similar biological functions , 2016, Journal of hepatology.

[9]  F. Ginhoux,et al.  Tissue-Resident Macrophage Ontogeny and Homeostasis. , 2016, Immunity.

[10]  F. Huang,et al.  Forced Activation of Notch in Macrophages Represses Tumor Growth by Upregulating miR-125a and Disabling Tumor-Associated Macrophages. , 2016, Cancer research.

[11]  Y. Saeys,et al.  Bone marrow-derived monocytes give rise to self-renewing and fully differentiated Kupffer cells , 2016, Nature Communications.

[12]  Carla M. T. Bauer,et al.  Self-renewing resident arterial macrophages arise from embryonic CX3CR1+ precursors and circulating monocytes immediately after birth , 2015, Nature Immunology.

[13]  Jing-quan Li,et al.  Targeting of tumour-infiltrating macrophages via CCL2/CCR2 signalling as a therapeutic strategy against hepatocellular carcinoma , 2015, Gut.

[14]  J. Carethers,et al.  Genetics and Genetic Biomarkers in Sporadic Colorectal Cancer. , 2015, Gastroenterology.

[15]  S. Monga.  β-Catenin Signaling and Roles in Liver Homeostasis, Injury, and Tumorigenesis. , 2015, Gastroenterology.

[16]  Sharad K Sharma,et al.  Pulmonary Alveolar Macrophages Contribute to the Premetastatic Niche by Suppressing Antitumor T Cell Responses in the Lungs , 2015, The Journal of Immunology.

[17]  F. Ginhoux,et al.  C-Myb(+) erythro-myeloid progenitor-derived fetal monocytes give rise to adult tissue-resident macrophages. , 2015, Immunity.

[18]  Runsheng Chen,et al.  The long noncoding RNA lncTCF7 promotes self-renewal of human liver cancer stem cells through activation of Wnt signaling. , 2015, Cell stem cell.

[19]  A. Jemal,et al.  Global cancer statistics, 2012 , 2015, CA: a cancer journal for clinicians.

[20]  Gérard Eberl,et al.  Liver-resident macrophage necroptosis orchestrates type 1 microbicidal inflammation and type-2-mediated tissue repair during bacterial infection. , 2015, Immunity.

[21]  K. Dou,et al.  Myeloid‐specific disruption of recombination signal binding protein Jκ ameliorates hepatic fibrosis by attenuating inflammation through cylindromatosis in mice , 2015, Hepatology.

[22]  F. Geissmann,et al.  Tissue-resident macrophages originate from yolk-sac-derived erythro-myeloid progenitors , 2014, Nature.

[23]  T. Welling,et al.  Tumor-associated macrophages produce interleukin 6 and signal via STAT3 to promote expansion of human hepatocellular carcinoma stem cells. , 2014, Gastroenterology.

[24]  Ming O. Li,et al.  The ontogeny of tumor-associated macrophages: a new understanding of cancer-elicited inflammation , 2014, Oncoimmunology.

[25]  Cassandra R Diegel,et al.  Beta‐catenin signaling in murine liver zonation and regeneration: A Wnt‐Wnt situation! , 2014, Hepatology.

[26]  H. Fiegl,et al.  In situ proliferation contributes to accumulation of tumor‐associated macrophages in spontaneous mammary tumors , 2014, European journal of immunology.

[27]  G. Randolph,et al.  Origin and functions of tissue macrophages. , 2014, Immunity.

[28]  Jeffrey W Pollard,et al.  Tumor-associated macrophages: from mechanisms to therapy. , 2014, Immunity.

[29]  Will Liao,et al.  The cellular and molecular origin of tumor-associated macrophages , 2014, Science.

[30]  N. Bird,et al.  Natural history of hepatic metastases from colorectal cancer--pathobiological pathways with clinical significance. , 2014, World journal of gastroenterology.

[31]  L. Coussens,et al.  CSF1R inhibition delays cervical and mammary tumor growth in murine models by attenuating the turnover of tumor-associated macrophages and enhancing infiltration by CD8+ T cells , 2013, Oncoimmunology.

[32]  D. Hume,et al.  IL-4 directly signals tissue-resident macrophages to proliferate beyond homeostatic levels controlled by CSF-1 , 2013, The Journal of experimental medicine.

[33]  E. Ager,et al.  Bimodal role of Kupffer cells during colorectal cancer liver metastasis , 2013, Cancer Biology & Therapy.

[34]  G. Trinchieri,et al.  The proinflammatory myeloid cell receptor TREM-1 controls Kupffer cell activation and development of hepatocellular carcinoma. , 2012, Cancer research.

[35]  D. Gabrilovich,et al.  Coordinated regulation of myeloid cells by tumours , 2012, Nature Reviews Immunology.

[36]  G. Evan,et al.  Role of c-MYC in alternative activation of human macrophages and tumor-associated macrophage biology. , 2012, Blood.

[37]  T. Luedde,et al.  Senescence surveillance of pre-malignant hepatocytes limits liver cancer development , 2011, Nature.

[38]  Chulan Kwon,et al.  Notch Post-Translationally Regulates β-Catenin Protein in Stem and Progenitor Cells , 2011, Nature Cell Biology.

[39]  L. Esserman,et al.  Proliferating macrophages associated with high grade, hormone receptor negative breast cancer and poor clinical outcome , 2011, Breast Cancer Research and Treatment.

[40]  Jinghang Zhang,et al.  CCL2 recruits inflammatory monocytes to facilitate breast tumor metastasis , 2011, Nature.

[41]  C. Nguyên,et al.  Inhibition of Wnt/β-catenin/CREB binding protein (CBP) signaling reverses pulmonary fibrosis , 2010, Proceedings of the National Academy of Sciences.

[42]  Xingbin Hu,et al.  Notch signaling determines the M1 versus M2 polarization of macrophages in antitumor immune responses. , 2010, Cancer research.

[43]  Jeffrey W. Pollard,et al.  Macrophage Diversity Enhances Tumor Progression and Metastasis , 2010, Cell.

[44]  Lieping Chen,et al.  Kupffer cell suppression of CD8+ T cells in human hepatocellular carcinoma is mediated by B7-H1/programmed death-1 interactions. , 2009, Cancer research.

[45]  Raphael Kopan,et al.  The Canonical Notch Signaling Pathway: Unfolding the Activation Mechanism , 2009, Cell.

[46]  P. Hainaut,et al.  Common dysregulation of Wnt/Frizzled receptor elements in human hepatocellular carcinoma , 2008, British Journal of Cancer.

[47]  A. Martinez Arias,et al.  Wnt/Notch signalling and information processing during development , 2008, Development.

[48]  M. Al-Fageeh,et al.  Cloning of the rat beta-catenin gene (Ctnnb1) promoter and its functional analysis compared with the Catnb and CTNNB1 promoters. , 2004, Genomics.

[49]  W. Reith,et al.  Conditional gene targeting in macrophages and granulocytes using LysMcre mice , 1999, Transgenic Research.

[50]  K. Movahedi,et al.  Tissue-resident versus monocyte-derived macrophages in the tumor microenvironment. , 2016, Biochimica et biophysica acta.

[51]  Michael Karin,et al.  References and Notes Supporting Online Material Materials and Methods Som Text Figs. S1 to S6 Tables S1 to S4 Gender Disparity in Liver Cancer Due to Sex Differences in Myd88-dependent Il-6 Production , 2022 .