S-Space College of Medicine/School of Medicine (의과대학/대학원) Dept. of Clinical Medical Sciences (임상의과학과) Theses (Ph.D. / Sc.D._임상의과학과)
Radiation-induced change of PD-1/PD-L1 immune checkpoint in mouse and human colorectal cancer models
대장직장암 모델 기반 방사선 조사 후 PD-1/PD-L1 면역 관문의 변화에 관한 연구
|dc.description||학위논문 (박사)-- 서울대학교 대학원 : 의과대학 임상의과학과, 2018. 2. 우홍균.||-|
|dc.description.abstract||“Nadir”, representing minimal tumor volume||-|
|dc.description.abstract||and “Regrowth”, with regrown tumors after RT. Defining the Day 1 as an initiation of RT, tumor tissues were obtained on Day 1, 6, 12, and 22, and Day 1, 6, 10, and 20, with single ablative and fractionated dose regimen, respectively. PD-L1 expression on tumor cells, PD-1 expression on tumor-infiltrating CD4+ and CD8+ T cells, and proportions of tumor-infiltrating CD4+ and CD8+ T cell populations were estimated using flow cytometry analysis. Considering human data, we conducted paired analysis using pre-CRT biopsies and the corresponding post-CRT resected tissues of 123 rectal cancer patients undergoing preoperative CRT followed by surgery between 2005 and 2012. Immunohistochemistry of PD-L1, PD-1, and CD8 was analyzed along with other clinicopathologic features and survival outcomes.
Results: PD-L1 expression on mouse tumor cells surged within a few days after completion of RT, followed by abrupt decreases on the “Nadir” and “Regrowth” phases (P < .001 and equal to .002 for single ablative and fractionated RT, respectively). PD-1-positivity (%) on CD4+ T cells was not significantly different according to different time points (P = .656 and .223 for single ablative and fractionated RT, respectively). On the contrary, PD-1-positive proportions (%) in CD8+ T cells sharply increased, and the high-level was sustained until the “Regrowth” phase (P < .001 for all paired comparisons between “Pre-RT” and others). During RT response of “Early” and “Nadir” time points, CD4+ T cells decreased, but CD8+ T cells increased. The alterations were reversed at “Regrowth” phase, with increasing and decreasing again in CD4+ and CD8+ T cell populations, respectively (P < .001 all paired comparisons between “Pre-RT” and others).
In the immunohistochemistry of rectal cancer, PD-L1 expression levels and density of CD8+ tumor-infiltrating lymphocytes (TILs) increased after CRT (P < .001 for both). Considering PD-1 expression, its pre-CRT intensity was scanty, but markedly increased after CRT. With cutoffs using each median value, sustained higher expression of PD-L1 at pre- and post-CRT (high-to-high) was associated with less increase in the density of CD8+ TILs (P = .020). Patients with sustained high-to-high PD-L1 expression had poorer overall survival (OS) and disease-free interval (DFI) in univariate Kaplan-Meier analysis (P = .018 and .029, respectively), with inferior DFI in low-to-low density CD8+ TILs (P = .010). In multivariate analysis, two subgroups with high baseline PD-L1 expression level showed worse OS, but the highest risk was observed with the high-to-high alteration (hazard ratio [HR] 8.34, 95% confidence interval [CI] 1.85–37.53 and HR 11.03, 95% CI 2.33–52.29 for high-to-low and high-to-high, respectively).
Conclusions: This study verified radiation-induced immunologic shift toward increases of the PD-1/PD-L1 checkpoint activity and density of CD8+ TILs. However, the change was maximal at the early phase of RT response, which highlights the need of concurrent combinatory strategy of PD-L1 blockade and RT. The alteration profiles of checkpoint-related molecules identified the subset of patients with poor prognosis, suggesting potential candidates who can benefit from combining checkpoint inhibitors.
|dc.description.abstract||Introduction: Recent progress in immunotherapy has introduced programmed death-1 (PD-1)/ programmed death-ligand 1 (PD-L1) blockade as a novel target to eradicate tumors, but its use has been mainly confined to recurrent or metastatic settings. Radiotherapy (RT), a major part of anti-cancer treatment, directly kills tumor cells, and subsequent anti-tumor immune responses are up-regulated. However, immunologic impacts of RT on PD-1/PD-L1 immune checkpoint activity has not been much investigated. This study evaluated RT-induced alterations of the PD-1/PD-L1 checkpoint molecules based on a murine colon carcinoma and human rectal cancer treated with preoperative chemoradiotherapy (CRT).
Methods: CT26 colon carcinoma cell line was subcutaneously inoculated on the right hind leg of BALB/c mice. Based on tumor growth curves after irradiation of 15 Gy x 1 fx or 5 Gy x 3 fx, mouse tumors were surgically resected on 4 different time points: “Pre-RT”, non-irradiated status just prior to initiation of RT
|dc.description.abstract||“Early”, the early phase of RT response||-|
Materials and methods 4
Cell line and animals 4
Preliminary experiments 4
Mouse tumor model 6
Single cell suspension of mouse tumors 8
Flow cytometry analysis 8
Patient population 11
Treatment of patients 11
Pathologic evaluation 13
Statistical analysis 14
Tumor growth curve after irradiation 16
PD-L1 expressions on tumor cells 17
PD-1 expression on CD4+ and CD8+ T cells 18
Proportions of CD4+ and CD8+ T cell subsets 20
Clinicopathologic characteristics of rectal cancer patients 23
Change of PD-L1, CD8+ and PD-1+ TILs before and after CRT 26
Survival analysis 37
국문 초록 61
|dc.subject||mouse tumor model||-|
|dc.title||Radiation-induced change of PD-1/PD-L1 immune checkpoint in mouse and human colorectal cancer models||-|
|dc.title.alternative||대장직장암 모델 기반 방사선 조사 후 PD-1/PD-L1 면역 관문의 변화에 관한 연구||-|
|dc.contributor.AlternativeAuthor||Yu Jin Lim||-|
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- College of Medicine/School of Medicine (의과대학/대학원)Dept. of Clinical Medical Sciences (임상의과학과)Theses (Ph.D. / Sc.D._임상의과학과)