Targeting CXCR4-dependent immunosuppressive Ly6Clow monocytes improves antiangiogenic therapy in colorectal cancer

Keehoon Jung, Takahiro Heishi, Joao Incio, Yuhui Huang, Elizabeth Y. Beech, Matthias Pinter, William W. Ho, Kosuke Kawaguchi, Nuh N. Rahbari, Euiheon Chung, Jun Ki Kim, Jeffrey W. Clark, Christopher G. Willett, Seok Hyun Yun, Andrew D. Luster, Timothy P. Padera, Rakesh K. Jain, Dai Fukumura

Research output: Contribution to journalArticle

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Abstract

Antiangiogenic therapy with antibodies against VEGF (bevacizumab) or VEGFR2 (ramucirumab) has been proven efficacious in colorectal cancer (CRC) patients. However, the improvement in overall survival is modest and only in combination with chemotherapy. Thus, there is an urgent need to identify potential underlying mechanisms of resistance specific to antiangiogenic therapy and develop strategies to overcome them. Here we found that anti-VEGFR2 therapy up-regulates both C-X-C chemokine ligand 12 (CXCL12) and C-X-C chemokine receptor 4 (CXCR4) in orthotopic murine CRC models, including SL4 and CT26. Blockade of CXCR4 signaling significantly enhanced treatment efficacy of anti-VEGFR2 treatment in both CRC models. CXCR4 was predominantly expressed in immunosuppressive innate immune cells, which are recruited to CRCs upon anti-VEGFR2 treatment. Blockade of CXCR4 abrogated the recruitment of these innate immune cells. Importantly, these myeloid cells were mostly Ly6Clow monocytes and not Ly6Chigh monocytes. To selectively deplete individual innate immune cell populations, we targeted key pathways in Ly6Clow monocytes (Cx3cr1/ mice), Ly6Chigh monocytes (CCR2/ mice), and neutrophils (anti-Ly6G antibody) in combination with CXCR4 blockade in SL4 CRCs. Depletion of Ly6Clow monocytes or neutrophils improved anti-VEGFR2–induced SL4 tumor growth delay similar to the CXCR4 blockade. In CT26 CRCs, highly resistant to anti-VEGFR2 therapy, CXCR4 blockade enhanced anti-VEGFR2–induced tumor growth delay but specific depletion of Ly6G+ neutrophils did not. The discovery of CXCR4-dependent recruitment of Ly6Clow monocytes in tumors unveiled a heretofore unknown mechanism of resistance to anti-VEGF therapies. Our findings also provide a rapidly translatable strategy to enhance the outcome of anti-VEGF cancer therapies.

Original languageEnglish
Pages (from-to)10455-10460
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume114
Issue number39
DOIs
StatePublished - 26 Sep 2017

Fingerprint

CXC Chemokines
Chemokine Receptors
Immunosuppressive Agents
Monocytes
Colorectal Neoplasms
Vascular Endothelial Growth Factor A
Neutrophils
Therapeutics
Neoplasms
CC Chemokines
Myeloid Cells
Growth
Combination Drug Therapy
Anti-Idiotypic Antibodies
Up-Regulation
Ligands
Survival

Keywords

  • Antiangiogenic therapy
  • CXCR4
  • Ly6C monocyte
  • Tumor immunity
  • Tumor microenvironment

Cite this

Jung, Keehoon ; Heishi, Takahiro ; Incio, Joao ; Huang, Yuhui ; Beech, Elizabeth Y. ; Pinter, Matthias ; Ho, William W. ; Kawaguchi, Kosuke ; Rahbari, Nuh N. ; Chung, Euiheon ; Kim, Jun Ki ; Clark, Jeffrey W. ; Willett, Christopher G. ; Yun, Seok Hyun ; Luster, Andrew D. ; Padera, Timothy P. ; Jain, Rakesh K. ; Fukumura, Dai. / Targeting CXCR4-dependent immunosuppressive Ly6Clow monocytes improves antiangiogenic therapy in colorectal cancer. In: Proceedings of the National Academy of Sciences of the United States of America. 2017 ; Vol. 114, No. 39. pp. 10455-10460.
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abstract = "Antiangiogenic therapy with antibodies against VEGF (bevacizumab) or VEGFR2 (ramucirumab) has been proven efficacious in colorectal cancer (CRC) patients. However, the improvement in overall survival is modest and only in combination with chemotherapy. Thus, there is an urgent need to identify potential underlying mechanisms of resistance specific to antiangiogenic therapy and develop strategies to overcome them. Here we found that anti-VEGFR2 therapy up-regulates both C-X-C chemokine ligand 12 (CXCL12) and C-X-C chemokine receptor 4 (CXCR4) in orthotopic murine CRC models, including SL4 and CT26. Blockade of CXCR4 signaling significantly enhanced treatment efficacy of anti-VEGFR2 treatment in both CRC models. CXCR4 was predominantly expressed in immunosuppressive innate immune cells, which are recruited to CRCs upon anti-VEGFR2 treatment. Blockade of CXCR4 abrogated the recruitment of these innate immune cells. Importantly, these myeloid cells were mostly Ly6Clow monocytes and not Ly6Chigh monocytes. To selectively deplete individual innate immune cell populations, we targeted key pathways in Ly6Clow monocytes (Cx3cr1−/− mice), Ly6Chigh monocytes (CCR2−/− mice), and neutrophils (anti-Ly6G antibody) in combination with CXCR4 blockade in SL4 CRCs. Depletion of Ly6Clow monocytes or neutrophils improved anti-VEGFR2–induced SL4 tumor growth delay similar to the CXCR4 blockade. In CT26 CRCs, highly resistant to anti-VEGFR2 therapy, CXCR4 blockade enhanced anti-VEGFR2–induced tumor growth delay but specific depletion of Ly6G+ neutrophils did not. The discovery of CXCR4-dependent recruitment of Ly6Clow monocytes in tumors unveiled a heretofore unknown mechanism of resistance to anti-VEGF therapies. Our findings also provide a rapidly translatable strategy to enhance the outcome of anti-VEGF cancer therapies.",
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Jung, K, Heishi, T, Incio, J, Huang, Y, Beech, EY, Pinter, M, Ho, WW, Kawaguchi, K, Rahbari, NN, Chung, E, Kim, JK, Clark, JW, Willett, CG, Yun, SH, Luster, AD, Padera, TP, Jain, RK & Fukumura, D 2017, 'Targeting CXCR4-dependent immunosuppressive Ly6Clow monocytes improves antiangiogenic therapy in colorectal cancer', Proceedings of the National Academy of Sciences of the United States of America, vol. 114, no. 39, pp. 10455-10460. https://doi.org/10.1073/pnas.1710754114

Targeting CXCR4-dependent immunosuppressive Ly6Clow monocytes improves antiangiogenic therapy in colorectal cancer. / Jung, Keehoon; Heishi, Takahiro; Incio, Joao; Huang, Yuhui; Beech, Elizabeth Y.; Pinter, Matthias; Ho, William W.; Kawaguchi, Kosuke; Rahbari, Nuh N.; Chung, Euiheon; Kim, Jun Ki; Clark, Jeffrey W.; Willett, Christopher G.; Yun, Seok Hyun; Luster, Andrew D.; Padera, Timothy P.; Jain, Rakesh K.; Fukumura, Dai.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 39, 26.09.2017, p. 10455-10460.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Targeting CXCR4-dependent immunosuppressive Ly6Clow monocytes improves antiangiogenic therapy in colorectal cancer

AU - Jung, Keehoon

AU - Heishi, Takahiro

AU - Incio, Joao

AU - Huang, Yuhui

AU - Beech, Elizabeth Y.

AU - Pinter, Matthias

AU - Ho, William W.

AU - Kawaguchi, Kosuke

AU - Rahbari, Nuh N.

AU - Chung, Euiheon

AU - Kim, Jun Ki

AU - Clark, Jeffrey W.

AU - Willett, Christopher G.

AU - Yun, Seok Hyun

AU - Luster, Andrew D.

AU - Padera, Timothy P.

AU - Jain, Rakesh K.

AU - Fukumura, Dai

PY - 2017/9/26

Y1 - 2017/9/26

N2 - Antiangiogenic therapy with antibodies against VEGF (bevacizumab) or VEGFR2 (ramucirumab) has been proven efficacious in colorectal cancer (CRC) patients. However, the improvement in overall survival is modest and only in combination with chemotherapy. Thus, there is an urgent need to identify potential underlying mechanisms of resistance specific to antiangiogenic therapy and develop strategies to overcome them. Here we found that anti-VEGFR2 therapy up-regulates both C-X-C chemokine ligand 12 (CXCL12) and C-X-C chemokine receptor 4 (CXCR4) in orthotopic murine CRC models, including SL4 and CT26. Blockade of CXCR4 signaling significantly enhanced treatment efficacy of anti-VEGFR2 treatment in both CRC models. CXCR4 was predominantly expressed in immunosuppressive innate immune cells, which are recruited to CRCs upon anti-VEGFR2 treatment. Blockade of CXCR4 abrogated the recruitment of these innate immune cells. Importantly, these myeloid cells were mostly Ly6Clow monocytes and not Ly6Chigh monocytes. To selectively deplete individual innate immune cell populations, we targeted key pathways in Ly6Clow monocytes (Cx3cr1−/− mice), Ly6Chigh monocytes (CCR2−/− mice), and neutrophils (anti-Ly6G antibody) in combination with CXCR4 blockade in SL4 CRCs. Depletion of Ly6Clow monocytes or neutrophils improved anti-VEGFR2–induced SL4 tumor growth delay similar to the CXCR4 blockade. In CT26 CRCs, highly resistant to anti-VEGFR2 therapy, CXCR4 blockade enhanced anti-VEGFR2–induced tumor growth delay but specific depletion of Ly6G+ neutrophils did not. The discovery of CXCR4-dependent recruitment of Ly6Clow monocytes in tumors unveiled a heretofore unknown mechanism of resistance to anti-VEGF therapies. Our findings also provide a rapidly translatable strategy to enhance the outcome of anti-VEGF cancer therapies.

AB - Antiangiogenic therapy with antibodies against VEGF (bevacizumab) or VEGFR2 (ramucirumab) has been proven efficacious in colorectal cancer (CRC) patients. However, the improvement in overall survival is modest and only in combination with chemotherapy. Thus, there is an urgent need to identify potential underlying mechanisms of resistance specific to antiangiogenic therapy and develop strategies to overcome them. Here we found that anti-VEGFR2 therapy up-regulates both C-X-C chemokine ligand 12 (CXCL12) and C-X-C chemokine receptor 4 (CXCR4) in orthotopic murine CRC models, including SL4 and CT26. Blockade of CXCR4 signaling significantly enhanced treatment efficacy of anti-VEGFR2 treatment in both CRC models. CXCR4 was predominantly expressed in immunosuppressive innate immune cells, which are recruited to CRCs upon anti-VEGFR2 treatment. Blockade of CXCR4 abrogated the recruitment of these innate immune cells. Importantly, these myeloid cells were mostly Ly6Clow monocytes and not Ly6Chigh monocytes. To selectively deplete individual innate immune cell populations, we targeted key pathways in Ly6Clow monocytes (Cx3cr1−/− mice), Ly6Chigh monocytes (CCR2−/− mice), and neutrophils (anti-Ly6G antibody) in combination with CXCR4 blockade in SL4 CRCs. Depletion of Ly6Clow monocytes or neutrophils improved anti-VEGFR2–induced SL4 tumor growth delay similar to the CXCR4 blockade. In CT26 CRCs, highly resistant to anti-VEGFR2 therapy, CXCR4 blockade enhanced anti-VEGFR2–induced tumor growth delay but specific depletion of Ly6G+ neutrophils did not. The discovery of CXCR4-dependent recruitment of Ly6Clow monocytes in tumors unveiled a heretofore unknown mechanism of resistance to anti-VEGF therapies. Our findings also provide a rapidly translatable strategy to enhance the outcome of anti-VEGF cancer therapies.

KW - Antiangiogenic therapy

KW - CXCR4

KW - Ly6C monocyte

KW - Tumor immunity

KW - Tumor microenvironment

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