title: Connecting the Dots: Therapy-Induced Senescence and a Tumor-Suppressive Immune Microenvironment.
abstract: BACKGROUND
  Tumor cell senescence is a common outcome of anticancer therapy. Here we investigated how therapy-induced senescence (TIS) affects tumor-infiltrating leukocytes (TILs) and the efficacy of immunotherapy in melanoma.
  
  
  METHODS
  Tumor senescence was induced by AURKA or CDK4/6 inhibitors (AURKAi, CDK4/6i). Transcriptomes of six mouse tumors with differential response to AURKAi were analyzed by RNA sequencing, and TILs were characterized by flow cytometry. Chemokine RNA and protein expression were determined by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Therapeutic response was queried in immunodeficient mice, in mice with CCL5-deficient tumors, and in mice cotreated with CD137 agonist to activate TILs. CCL5 expression in reference to TIS and markers of TILs was studied in human melanoma tumors using patient-derived xenografts (n = 3 patients, n = 3 mice each), in AURKAi clinical trial samples (n = 3 patients, before/after therapy), and in The Cancer Genome Atlas (n = 278). All statistical tests were two-sided.
  
  
  RESULTS
  AURKAi response was associated with induction of the immune transcriptome (P = 3.5 x 10-29) while resistance inversely correlated with TIL numbers (Spearman r = -0.87, P < .001). AURKAi and CDK4/6i promoted the recruitment of TILs by inducing CCL5 secretion in melanoma cells (P ≤ .005) in an NF-κB-dependent manner. Therapeutic response to AURKAi was impaired in immunodeficient compared with immunocompetent mice (0% vs 67% tumors regressed, P = .01) and in mice bearing CCL5-deficient vs control tumors (P = .61 vs P = .02); however, AURKAi response was greatly enhanced in mice also receiving T-cell-activating immunotherapy (P < .001). In human tumors, CCL5 expression was also induced by AURKAi (P ≤ .02) and CDK4/6i (P = .01) and was associated with increased immune marker expression (P = 1.40 x 10-93).
  
  
  CONCLUSIONS
  Senescent melanoma cells secret CCL5, which promotes recruitment of TILs. Combining TIS with immunotherapy that enhances tumor cell killing by TILs is a promising novel approach to improve melanoma outcomes.
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE). anna.e.vilgelm@vanderbilt.edu.
authors.forename: Anna E
authors.initials: AE
authors.lastname: Vilgelm
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: C Andrew
authors.initials: CA
authors.lastname: Johnson
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Nripesh
authors.initials: N
authors.lastname: Prasad
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Jinming
authors.initials: J
authors.lastname: Yang
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Sheau-Chiann
authors.initials: SC
authors.lastname: Chen
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Ayers
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Jeff S
authors.initials: JS
authors.lastname: Pawlikowski
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Dayanidhi
authors.initials: D
authors.lastname: Raman
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Sosman
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Mark
authors.initials: M
authors.lastname: Kelley
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Ecsedy
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Yu
authors.initials: Y
authors.lastname: Shyr
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Shawn E
authors.initials: SE
authors.lastname: Levy
authors.affiliation: Tennessee Valley Healthcare System, Department of Veterans Affairs, Nashville, TN (AEV, CAJ, JY, JSP, DR, AR); Department of Cancer Biology (AEV, AJ, JY, SCC, JSP, DR, AR), Center for Quantitative Sciences (SCC), Division of Cancer Biostatistics, Department of Biostatistics (GDA, YS), Division of Hematology/Oncology, Department of Medicine (JAS), Vanderbilt University Medical Center, Nashville, TN; HudsonAlpha Institute for Biotechnology, Huntsville, Alabama (NP, SEL); Division of Surgical Oncology, Department of Surgery, Vanderbilt University School of Medicine, Nashville, TN (MK); Translational Medicine, Takeda Pharmaceuticals International Co, Cambridge, MA (JAE).
authors.forename: Ann
authors.initials: A
authors.lastname: Richmond
mesh_terms: D000818:Animals; D064096:Aurora Kinase A / Q000037:antagonists & inhibitors; D045744:Cell Line, Tumor; D016922:Cellular Senescence; D018946:Chemokine CCL5 / Q000378:metabolism; D051358:Cyclin-Dependent Kinase 4 / Q000037:antagonists & inhibitors; D051361:Cyclin-Dependent Kinase 6 / Q000037:antagonists & inhibitors; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D064593:Heterografts; D006801:Humans; D007121:Immunocompetence; D016867:Immunocompromised Host; D007167:Immunotherapy / Q000379:methods*; D008213:Lymphocyte Activation / Q000276:immunology; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008545:Melanoma / Q000276:immunology / Q000473:pathology* / Q000628:therapy*; D051379:Mice; D060888:Real-Time Polymerase Chain Reaction; D012878:Skin Neoplasms / Q000276:immunology / Q000473:pathology* / Q000628:therapy*; D059467:Transcriptome; D059016:Tumor Microenvironment / Q000276:immunology*; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000276:immunology
chemical_list: C483024:CCL5 protein, human; C503460:Ccl5 protein, mouse; D018946:Chemokine CCL5; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; C576917:AURKA protein, human; D064096:Aurora Kinase A; C495900:CDK4 protein, human; C495911:CDK6 protein, human; D051358:Cyclin-Dependent Kinase 4; D051361:Cyclin-Dependent Kinase 6