Istituto Nazionale Tumori di Napoli, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) "G. Pascale", Naples, Italy.
Oncohaematology Department, Azienda Ospedaliera di Rilievo Nazionale (A.O.R.N.) Santobono-Pausilipon di Napoli, Naples, Italy.
Front Immunol. 2022 Jul 22;13:898561. doi: 10.3389/fimmu.2022.898561. eCollection 2022.
We previously reported rare regressive genetic trajectories of pathogenic mutations as a specific hallmark of the genuine oligometastatic status in colorectal cancer (CRC).
Survival and prognostic impact of disease extent in 140 metastatic CRC patients were evaluated through the Kaplan-Meyer curves and the Log-Rank test. mutations were assessed through the Illumina NovaSeq 6000 platform and TruSight™ Oncology 500 kit. HLA typing was carried out by PCR with sequence-specific oligonucleotides. Lymphocyte densities in tumors were expressed as cells per square millimeter. NKs isolated and CD8 from NK-depleted PBMCs were characterized through flow cytometry. CD107a externalization was evaluated as NKs/CD8 cytotoxicity toward human colon cancer cells HT29, SW620, HCT116, and LS174T carrying different mutations.
The oligometastatic status was a strong and independent variable for survival (HR: 0.08 vs. polymetastatic disease; 95% CI: 0.02-0.26; < 0.001). Eighteen oligometastatic patients were selected. Twelve were alive at the last follow-up, and 9 were characterized. Genetic regression of was observed in 3 patients: patient (PAT)2, PAT5, and PAT8. PAT2 and PAT5 presented the highest levels of GrzB lymphocytes in the tumor cores of the metastases (120 ± 11.2 and 132 ± 12.2 cells/mm, respectively). Six out of 9 patients (67%), including PAT2 and PAT5, expressed HLA-C7. Twopatients (PAT2 and PAT5) presented high CD3/CD8-dependent cytotoxicity against HLA-C7+ SW620 cells (p.G12V-mutated cells), which was consistent with their observed mutational regression (p.G12V/p.G13D in primary→p.G13D in metastatic tumor).
We provide evidence that CD3/CD8 lymphocytes from oligometastatic CRC patients display differential cytotoxicity against human colon cancer cells carrying mutations. This could provide an interesting basis for monitoring oligometastatic disease and developing future adoptive immunotherapies.
我们之前报道过，致病性突变的罕见退行性遗传轨迹是结直肠癌（CRC）真正寡转移状态的特定标志。
通过 Kaplan-Meier 曲线和 Log-Rank 检验评估 140 例转移性 CRC 患者的疾病范围的生存和预后影响。通过 Illumina NovaSeq 6000 平台和 TruSight™ Oncology 500 试剂盒评估 突变。通过聚合酶链反应（PCR）与序列特异性寡核苷酸进行 HLA 分型。肿瘤中的淋巴细胞密度表示为每平方毫米的细胞数。分离的 NK 细胞和 NK 耗尽的 PBMC 中的 CD8 通过流式细胞术进行表征。CD107a 外化被评估为 NKs/CD8 对携带不同 突变的人结肠癌细胞 HT29、SW620、HCT116 和 LS174T 的细胞毒性。
寡转移状态是生存的强独立变量（HR：0.08 vs. 多转移疾病；95%CI：0.02-0.26；<0.001）。选择了 18 例寡转移患者。18 例患者在最后一次随访时存活，其中 9 例进行了特征分析。在 3 例患者中观察到 的遗传回归：患者（PAT）2、PAT5 和 PAT8。PAT2 和 PAT5 在转移灶肿瘤核心中的 GrzB 淋巴细胞水平最高（分别为 120±11.2 和 132±12.2 个/毫米）。9 例患者中的 6 例（67%），包括 PAT2 和 PAT5，表达 HLA-C7。2 例患者（PAT2 和 PAT5）对 HLA-C7+SW620 细胞（p.G12V 突变细胞）表现出高 CD3/CD8 依赖性细胞毒性，这与他们观察到的突变回归一致（p.G12V/p.G13D 在原发性→p.G13D 在转移性肿瘤）。
我们提供的证据表明，来自寡转移 CRC 患者的 CD3/CD8 淋巴细胞对携带 突变的人结肠癌细胞表现出不同的细胞毒性。这为监测寡转移疾病和开发未来的过继免疫疗法提供了有趣的基础。
