Tumor associated macrophages in endocrine-related cancers. Sun Wook Cho, M.D, Ph.D Seoul National University Hospital
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1 Tumor associated macrophages in endocrine-related cancers Sun Wook Cho, M.D, Ph.D Seoul National University Hospital
2 Tumor Microenvironment Fibroblasts Immune cells Angiogenesis Tumor microenvironment=cancer cells + stroma Heterogeneous and complex
3 Cancer Immunology Immunosurveillance Necrotic Cell debris Dendritic cell T cell NK T cell Immune evasion Macrophage Immunoediting
4 Cancer Immunology Classical immune actions Mature dendritic cell Immature dendritic cell Macropahge MDSC Immune Therapy CD8 T cell Finn, NEJM. 28 Th1 CD4 T cell Treg cell Th2 CD4 T cell Alternative immune modulations Host defense mechanism
5 Tumor-associated macrophage, TAM Classically activated TNF, IFNγ Microbial LPS Monocytes Alternative activated IL-4, IL-1 M-CSF inos Arginase I M1-macrophage M2-macrophage IL-12, IL-23, TNF, IL-1, CXCL1 Microbicidal Tumoricidal IL-1, IL-13, IL-4, CCL22 scavenger, mannose receptor Cell clearance/wound healing Tumor promotion
6 Interactions between macrophage and tumor cells M2-macrophage (TAM) B cell TReg cell T2h cell Promote tumor survival
7 Tumor cell Macrophage Cell growth Cell cycle Other mechanisms Protease, Adhesion molecule ( MMP-1, TIMP-1, ICAM-1) Signal transduction (IL-6) Inflammation (CSF-1, IL-6, IL-8, PDGF) Angiogenesis (CSF-1, IL-6, IL-8)
8 The relationship between macrophage densities and 5-yr survival rates of human cancers 5-year overall survival (%) Average CD163 positive area (%) Thyroid (PTC) Endometrium Kidney Prostate Stomach Breast Ovary Cervix Liver Esophagus Larynx GB Lung Pancreas Thyroid (ATC) CD yr OS NA
9 TAM densities in thyroid cancers P/D, CD68 P/D, CD163 M/D, CD68 W/D, CD68 High density of TAMs in PDTC correlates with invasion and decreased cancer-related survival. Ryder et al. Endocr Relat Cancer. 28
10 TAM densities in thyroid tissues Normal Adenoma PTC ATC Macrophage Normal (N=7) Adenoma (n=57) PTC (n=148) ATC (n=17) P-value Negative 6 (86 %) <.1 Low density 1 (14 %) 41 (72 %) 78 (57 %) 3 (18 %) High density 16 (28 %) 58 (43 %) 14 (82 %)
11 TAM densities in Papillary thyroid cancers TAMs were scored by the number of CD68+ cells/total tumor cells Low (<25%) High (25% to 7%) Kim, Cho et al. EnM 213
12 Clinical characteristics according to the TAM densities TAM density Low (n=79) CD163 High (n=64) P-value Sex (M/F) 13/66 12/52.72 Age of diagnosis 45.3 ± ± Tumor size (cm) 2.3 ± ± Extrathyroidal invasion 15 (22%) 13 (22%).997 LN metastasis 38 (62%) 48 (8%).31 Recurrence or metastasis 5 (16%) 9 (23%).429 CD31 + cell (%) 3 (5%) 11 (19%).16 CD68 + cell (%) 3 (5%) 25 (4%) <.1
13 Macrophages support tumor growth Tumor cell ±THP-1 injection 7 35 (days) Sac BHP1-3 only BHP1-3/THP-1 Tumor volume (mm 2 ) THP-1(+)/PBS THP-1(-)/PBS Days after implantation
14 Macrophages support tumor growth Tumor cell ±THP-1 injection 7 35 (days) Sac H&E BHP1-3 only BHP1-3 / THP-1 Area of necrosis (µm 2 ) BHP1-3 only BHP1-3 / THP-1 Area of necrosis
15 Clodronate liposome inhibited macrophage-related tumor growth Tumor cell ±THP-1 injection Clodronate liposome Peritumoral injection (3 times/ week) 7 35 (days) Sac Clodronate liposome 7 Tumor volume (mm 2 ) # # THP-1(+)/PBS THP-1(+)/Clodronate THP-1(-)/PBS THP-1(-)/Clodronate Days after implantation
16 Clodronate liposome inhibited macrophage-related tumor growth Tumor cell ±THP-1 injection Clodronate liposome Peritumoral injection (3 times/ week) 7 35 (days) Sac + THP-1(+)/PBS + THP-1(+)/Clodronate THP-1(-)/PBS THP-1(+)/PBS THP-1(+)/Clodronate Frequency Intensity of CD68 CD68+ (%) 3 2 1, # Day 1 15 Day 2 25, #
17 TAMs support tumor cell migrations Wound healing assay Transwell assay 3D invasion assay + macrophage
18 TAM increased angiogenesis in tumor microenvironment Control CM BHP1-3 CM Control CM BHP1-3 CM + macrophage Total tube length (µm/field) THP-1 (-) THP-1 (+)
19 Tumor cell Macrophage Tumor growth How it works? Tumor cell migration Angiogenesis
20 CXCL16 expressions in tumor/macrophage co-cultures H.Tori (normal thyroid cells) BHP1-3, KTC-1 (PTC cells) + macrophage Conditioned medium (CM) + macrophage Normal thyrocyte KTC-1 BHP1-3 CXCL16 (pg/ml) H tori BHP1-3 KTC-1 macrophage H tori BHP1-3 KTC-1 +macrophage
21 CXCL16/CXCR6: a regulator of cancer metastasis Transmembrane CXCL16: growth Soluble CXCL16: metastasis
22 CXCL16 expressions in PTCs Control CM M/BHP1-3 CM Control CM Monocyte CM BHP1-3 CM M/BHP1-3 CM BHP1-3SC Macrophage CXCR6/GAPDH Monocyte 1 BHP1-3SC 2 # CXCL16/GAPDH # Monocyte 1 BHP1-3SC 2 CD163 + cell (No./1 3 µm 2 ) γ 2 =.28, P<.1 CXCR6 expressions in both tumor cell and stroma CXCR6 + cell (No./1 3 µm 2 )
23 CXCL16 mediated macrophage effects on tumor cell migration Vehicle CXCL16 THP-1 (-) THP-1 (+) Vehicle anti-cxcl16 TAPI-2 Migration length δ # 1 2 THP-1 (-) THP-1 (+) # mtor pakt AKT VEH CXCL16 VEH THP-1 (+) Anti- CXCL16 Densitometry THP-1(-) / VEH THP-1(-) / CXCL16 THP-1(+) / VEH THP-1(+) / anti-cxcl16 # # GAPDH (.) mtor/gapdh 1 pakt/akt 2
24 CXCL16 mediated macrophage effects on angiogenesis CM Vehicle CXCL16 Control 8 Control CM M/BHP1-3 CM M/BHP1-3 Vehicle anti-cxcl16 Total tube length (µm/field) #
25 ? Tumor cell Macrophage CXCL16/CXCR6 signaling Tumor growth Tumor cell migration Angiogenesis
26 Tumor recruit circulating monocytes and switch into M2- macrophages NOG mouse Bone marrow Tumor Tumor cell injection PKH26-labelled monocyte injection Frequency Intensity of red fluorescence (PKH26) (days) Sac PKH+ cell (%) in 1 6 cells Peripheral blood Bone marrow 6.3 ±.7 Tumor 13.3 ± 3.7
27 Tumor recruit circulating monocytes and switch into M2- macrophages NOG mouse mrna expressions in PKH26+ cell Bone marrow Tumor Tumor cell injection PKH26-labelled monocyte injection (days) Sac.5. CD163 IL-4 IL-12 TNFα
28 CD163 IL-1 CD26 Mock Relative gene expression Relative gene expression CM CM CM Relative gene expression
29 CD163 IL-1 CD Mock Anti-CXCL16 Relative gene expression Relative gene expression CM # # CM CM # Relative gene expression CD163 IL-1 CD26 IL-β inos Relative mrna expressions (rhcxcl16/vehicle) IL-1β Relative gene expression Relative gene expression. CM CM inos M-CSF (-) M-CSF (+).
30 TAM, as a therapeutic target in cancer therapy Blocking differentiation CSF-1, cfms inhibitors CCL2, monocloncal Ab (MLN122) Repolarizations Pantoprazole IL-12 Killing TAM in the tumor microenvironments Clodronate-loaded liposomes (CLIP) Zoledronic acid
31 Conclusions TAMs support tumor growth, invasion, and metastasis. CXCL16 signaling mediated TAM actions in human papillary thyroid cancers. TAM might be a potential therapeutic targets of human cancers.
32 Acknowledgements Seoul National University Pf. Young Joo Park Pf. Kyong Soo Park Pf. Do Joon Park Pf. Young A Kim Gachon University Pf. Byung-Chul Oh Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI) Young Investigater s award from Korean Endocrine society ( 향설연구비 )
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