肿瘤突变负荷对PD-1/PD-L1抑制剂治疗非小细胞肺癌临床疗效预测的Meta分析

目的 探讨肿瘤突变负荷(TMB)与PD-1/PD-L1抑制剂治疗非小细胞肺癌(NSCLC)疗效的相关性.方法 系统检索PubMed、Embase和Cochrane Library、CNKI、中国生物医学数据库(Chinese Biomedical Literature Database,CBM)和万方数据库,检索日期截...     

Correlation of PD-L1 Expression with Tumor Mutation Burden and Gene Signatures for Prognosis in Early-Stage Squamous Cell Lung Carcinoma

Objectives

Anti–programmed cell death 1 (PD-1)/programmed death ligand 1 (PD-L1) immunotherapy has demonstrated success in the treatment of advanced NSCLC. Recently, PD-1/PD-L1 blockade also has demonstrated interesting results in small trials of neoadjuvant treatment in stage IB to IIIA NSCLC. In addition, several clinical trials using anti–PD-1/PD-L1 immunotherapy as an adjuvant or neoadjuvant treatment in patients with resectable stage NSCLC are ongoing. However, few analyses of anti–PD-1/PD-L1 immunotherapy–related biomarkers in early-stage squamous cell lung carcinoma (SqCLC) have been reported. In this study, we evaluated PD-L1 protein expression, tumor mutation burden, and expression of an immune gene signature in early-stage SqCLC, providing data for identifying the potential role for patients with anti–PD-1/PD-L1 treatment in early-stage SqCLC.

Comparison of Three Sequencing Panels Used for the Assessment of Tumor Mutational Burden in NSCLC Reveals Low Comparability

IntroductionTumor mutational burden (TMB) has been proposed as a novel predictive biomarker for the stratification of patients with NSCLC undergoing immune checkpoint inhibitor (ICI) treatment. The assessment of TMB has recently been established using large targeted sequencing panels, and numerous studies are ongoing to harmonize TMB assessment. “Correlation” or the coefficient of determination has generally been used to evaluate the association between different panels. We hypothesized that these metrics might overestimate the comparability, especially for lower TMB values.MethodsA total of 30 samples from patients with NSCLC undergoing ICI treatment were consecutively sequenced using the following three large, targeted sequencing panels: FoundationOne, Oncomine TML, and QiaSeq TMB. The TMB values were compared in the whole patient population and in a subset of patients in which the TMB assessed by FoundationOne was between 5 and 25 mutations/Mb. Prediction of durable clinical benefit (>6 mo with no progression) was assessed using receiver operator characteristics, and optimal cutoff values were calculated using the Youden J statistic.ResultsCorrelation between the three targeted sequencing panels was strong in the whole patient population (R² > 0.79) but was dramatically reduced in the subset of patients with TMB of 5 to 25 mutations/Mb. The agreement assessed using the Bland-Altman method was also very low. All panels were able to predict durable clinical benefit in the TMB-high population.ConclusionsAssessment of TMB using the three targeted sequencing panels was possible and predictive of response to ICI treatment, but correlation was an inappropriate measurement to assess the association between the respective panels.     

Preliminary Evaluation of the New Tumor Marker, CYFRA 21-1, in Lung Cancer Patients

Serum samples from 137 lung cancer patients were examined byRIA to evaluate the clinical efficacy of the new tumor marker,CYFRA 21-1, which could identify the soluble fragment of cytokeratin19. The cut-off value was determined to be 2.2 ng/ml accordingto the receiver operating characteristic curve. The sensitivity,specificity and accuracy of the RIA for CYFRA 21-1 were 57.7,91.9 and 64.9%, respectively. The serum concentration of CYFRA21-1 and the sensitivity of the assay increased as the diseaseprogressed. Histologically, the sensitivity was highest forsquamous cell carcinomas (SQ) (76.5%) in comparison with adenocarcinomas(47.8%) and small cell lung cancers (42.1%) (P<0.01, P<0.05,respectively). The sensitivities for SQ were 60.0, 83.3, 80.0and 100% at stages I, II, III and IV, respectively. When comparedwith CEA (45.3%) and squamous cell carcinoma related antigen(SCC) (22.6%) in all lung carcinomas, CYFRA 21-1 showed thehighest sensitivity (57.7%), (P<0.05, P<0.01, respectively).In SQ, the sensitivity of the CYFRA 21-1 RIA was significantlyhigher than that of the assay for SCC (47.1%) (P<0.05). Inpatients with adenocarcinomas, the sensitivity of the CYFRA21-1 assay was almost the same as that for CEA (49.3%). In acombination of CYFRA 21-1 and CEA for non-small cell lung cancers(NSCLQ, the sensitivity and accuracy increased to 75.4 and 78.1%,respectively, although the specificity decreased to 86.5%. Itis concluded that CYFRA 21-1 could replace SCC, a less satisfactorytumor marker, for SQ of the lung. The potentiality of the combinationof CYFRA 21-1 and CEA for NSCLC should be estimated using largersamples in the near future     

非小细胞肺癌患者血清CYFRA_(21-1)和CEA的临床应用

 目的　研究癌标CYFRA2 1 1及CEA对非小细胞肺癌 (NSCLC)诊断、病情及预后的临床应用价值。方法　用放射免疫法检测 10 1例NSCLC患者和 112例良性肺疾病患者血清CYFRA2 1 1和CEA ,比较NSCLC的不同组织类型、临床分期、手术前后差异。结果　CYFRA2 1 1对NSCLC诊断敏感性、特异性、有效性为 72 .4 %、87.3%、79.1% ,CYFRA2 1 1对鳞癌诊断敏感性、有效性高于腺癌 (P <0 .0 5 ) ,而CEA对NSCLC诊断敏感性、特异性、有效性为 4 7.5 %、94 .0 %、71.6 % ,CEA对腺癌诊断敏感性、有效性高于鳞癌 (P <0 .0 1) ,CYFRA2 1 1与CEA联合应用可提高对NSCLC诊断敏感性、有效性。血清CYFRA2 1 1水平与TNM分期有关 ,随病情变化而改变。结论　血清CYFRA2 1 1检测特别是与CEA联合应用对NSCLC患者诊断、病情监测、预后判断有较高临床应用价值。     

Liver Kinase B1 (LKB1) in the pathogenesis of epithelial cancers

LKB1 acts as a master kinase, with its major protein targets being the family of AMPKs. Through activation of multiple signaling pathways, LKB1’s main physiologic functions involve regulating cellular growth, metabolism, and polarity. Germline mutations in LKB1 result in Peutz-Jeghers Syndrome, a rare cancer susceptibility syndrome. In addition, multiple LKB1 mutations have been identified in sporadic cancers, especially those of the lung. Recent studies from a variety of murine models have helped characterize LKB1’s role in the pathogenesis of epithelial cancers. In some tumor types, LKB1 might function chiefly to suppress cell growth or invasion, while in other cases, it may serve to prevent metastasis. Moreover, molecular signatures of individual tumors likely influence LKB1’s operational role, as multiple studies have shown that LKB1 can synergize with other tumor suppressors and/or oncogenes to accelerate tumorigenesis. To date, LKB1 has been considered mainly a tumor suppressor; however, some studies have suggested its potential oncogenic role, mainly through the suppression of apoptosis. In short, LKB1 is a tissue and context-specific kinase. This review aims to summarize our current understanding of its role in the pathogenesis of epithelial cancers.     

Urinary Bladder Lesions after the Chernobyl Accident: Immunohistochemical Assessment of p53, Proliferating Cell Nuclear Antigen, Cyclin D1 and p21WAF1/Cip1

During the 11-year period subsequent to the Chernobyl accident, the incidence of urinary bladder cancer in Ukraine has increased from 26.2 to 36.1 per 100,000 population. Cesium-137 (¹³⁷Cs) accounts for 80–90% of the incorporated radioactivity in this population, which has been exposed to long-term, low-dose ionizing radiation, and 80% of the more labile pool of cesium is excreted via the urine. The present study was performed to evaluate the histopathological features and the immunohistochemical status of p53, p21^WAF1/Cip1, cyclin D1 and PCNA (proliferating cell nuclear antigen) in urinary bladder mucos a of 55 males (49-92 years old) with benign prostatic hyperplasia who underwent surgery in Kiev, Ukraine, in 1995 and 1996. Group I (28 patients) inhabiting radiocontaminated areas of the country, group II (17 patients) from Kiev city with less radiocontamination and a control group III (10 patients) living in so-called "clean" areas of Ukraine were compared. In groups I and II, an increase in multiple areas of moderate or severe dysplasia or carcinoma in situ was seen in 42 (93%) of 45 cases. In addi tion, two small transitional cell carcinomas were found in one patient in each of groups I and II. Nuclear accumulation of p53, PCNA, cyclin D1, and to a lesser extent p21^WAF1/Cip1, was significantly increased in both groups I and II as compared with the control group III, indicating possible transformation events or enhancement of repair activities, that may precede the defect in the regulatory pathway itself, at least in the G1 phase of the cell cycle. Our results suggest that early malignant transformation is taking place in the bladder urothelium of people in the radiocontaminated areas of Ukraine and that this could possibly lead sometime in the future to an increased incidence of urinary bladder cancer.     

Suppression of Anti-tumor CD4+ T Cell Responsiveness in the Tumor-bearing State and Its Recovery in in vitro Culture Free of Tumor Burden

We investigated whether the responsiveness of anti-tumor CD4⁺ T cells suppressed in the tumor-bearing state is reversed in conditions free of tumor burden. Spleen cells from BALB/c mice bearing a syngeneic tumor (CSA1M) 1–3 wk after inoculation with CSA1M cells produced interleukin-2 (IL-2) and IL-4 upon in vitro culture without addition of exogenous tumor antigens. This lymphokine production was achieved through collaboration between anti-CSA1M CD4⁺ T cells and antigen-presenting cells (APC) that had been pulsed with CSA1M tumor antigens in vivo in the tumor-bearing state. However, spleen cells from late (8–10 wk) tumor-bearing stages produced reduced levels of lymphokine production despite the presence of comparable proportions of CD4⁺ T cells. Because APC in these cell populations exhibited enhanced capacities to present tumor antigens, reduced responsiveness was ascribed to the dysfunction of CD4⁺ T cells themselves. When spleen cells from early tumor-bearing mice were preincubated for 1–2 days and recultured in fresh medium, the magnitude of lymphokine production by these cells was not changed. In contrast, the same protocol of preincubation and reculture for cells from late tumor-bearing mice resulted in the recovery of anti-tumor lymphokine-producing capacity. The recovered capacity was comparable to or slightly higher than that expressed by cells from early tumor-bearing stages. Since the CD4⁺ T cell content did not significantly differ before and after preincubation, enhanced lymphokine production was due to the recovered responsiveness of anti-tumor CD4⁺ helper T cells. The recovery of anti-tumor responsiveness was also induced in vivo by tumor removal at the late tumor-bearing stage: spleen cells from mice 2–4 wk after tumor resection efficiently produced IL-2 and IL-4. These results indicate that the immunodysfunction of anti-tumor CD4⁺ T cells induced in the tumor-bearing state is reversible because release from tumor burden either by preincubation in vitro or by tumor removal in vivo results in almost complete recovery of the potent anti-tumor responsiveness initially expressed.     

Somatic Mutations of the PTEN/MMAC1 Gene in Fifteen Japanese Endometrial Cancers: Evidence for Inactivation of Both Alleles

Loss of heterozygosity (LOH) of chromosome 10q is observed in approximately 40% of endometrial cancers. Mutations in PTEN/MMAC1 , a gene recently isolated from the 10q23 region, are responsible for two dominantly inherited neoplastic syndromes, Cowden disease and Bannayan-Zonana syndrome. Somatic mutations of this gene have also been detected in sporadic cancers of the brain, prostate and breast. To investigate the potential role of this putative tumor suppressor gene in endometrial carcinogenesis as well, we examined 46 primary endometrial cancers for LOH at the 10q23 region, and for mutations in the entire coding region and exon-intron boundaries of the PTEN/MMAC1 gene. LOH was identified in half of the 38 informative cases, and subtle somatic mutations were detected in 15 tumors (33%). Our results suggest that of the genes studied so far in endometrial carcinomas, PTEN/MMAC1 is the most commonly mutated one, and that inactivation of both copies by allelic loss and/or mutation, a pattern that defines genes as "tumor suppressors,'contributes to tumorigenesis in endometrial cancers.     

Close Correlation of 1-β-D-Arabinofuranosylcytosine 5'-Triphosphate, an Intracellular Active Metabolite, to the Therapeutic Efficacy of N4-Behenoyl-1-β-D-arabinofuranosylcytosine Therapy for Acute Myelogenous Leukemia

N ⁴-Behenoyl-l-β-D-arabinofuranosylcytosine (BHAC), a prodrug of 1-β -D-arabinofuranosylcy-tosine, is used effectively for the treatment of leukemia in Japan. BHAC therapy may be more effective if it is delivered in conjunction with monitoring of 1-β -D-arabinofuranosylcytosine 5'-tri-phosphate (ara-CTP), the intracellular active metabolite of ara-C derived from BHAC. However, previous monitoring methods for ara-CTP were insufficiently sensitive. Here, using our new sensitive method, we evaluated the ara-CTP pharmacokinetics in relation to the therapeutic response in 11 acute myelogenous leukemia patients who received a 2-h infusion of BHAC (70 mg/m²) in combination remission induction therapy. ara-CTP could be monitored at levels under 1 μ M. BHAC maintained effective levels of plasma ara-C and intracellular ara-CTP for a longer time, even compared with historical values of high-dose ara-C. The area under the concentration-time curve of ara-CTP was significantly greater in the patients with complete remission than in the patients without response. This greater amount of ara-CTP was attributed to the higher ara-CTP concentrations achieved in the responding patients. There was no apparent difference of plasma ara-C pharmacokinetics between the two groups. Thus, for the first tune, the ara-CTP pharmacokinetics was evaluated in relation to the therapeutic effect of BHAC, and the importance of ara-CTP was proven. Administration of optimal BHAC therapy may require monitoring of the ara-CTP pharmacokinetics in each individual patient.     

Aberrant methylation of the RIZ1 gene in myelodysplastic syndrome and acute myeloid leukemia

We performed methylation specific PCR analysis on the RIZ1 promoter in MDS and AML. Methylation was detected in 17 of 34 MDS (50%) and 22 of 72 AML (31%) (p = 0.053). Methylation was detected in eleven of 17 secondary AML from MDS (65%), and eleven of 55 de novo AML (20%) (p = 0.0005). Bisulfite sequence revealed methylation at many CpG sites in the promoter. Decreased RIZ1 expression was accompanied by methylation in six of nine samples examined, while it was also observed in seven of 13 without methylation. Treatment of AML cells, that have RIZ1 methylation, with 5-Aza-dC, induced growth suppression with RIZ1 restoration. Our results suggest that the RIZ1 gene is inactivated in MDS and AML in part by methylation, whereas another mechanism should be involved in others.     

Expression of p21waf-1/cip-1 Is Significantly Induced in the Livers of LEC Rats with Chronic Liver Injury

It is reported that hepatocytes isolated from LEC rats with chronic liver injury show reduced growth activity in primary culture. To elucidate the molecular basis of this phenomenon, we examined expression of p21^waf-1/cip-1 and p27, cyclin-dependent kinase inhibitors, by northern blot analysis. The expression of p21^waf-1/cip-1 in the LEC rat liver was 3-fold higher than that of age-matched SD rat liver, while there was no significant difference in p27 expression level. Western blot analysis also revealed a significant increase in p2l^waf-1/cip-1 in the nuclear matrix fraction of the LEC rat liver. Immunohisto-chemically, p21^waf-1/cip-1 was detected in the nuclei of normal LEC rat hepatocytes, but not in those of hepatocellular carcinoma cells, suggesting selective growth of neoplastic hepatocytes.     

EBUS-TBNA as a Promising Method for the Evaluation of Tumor PD-L1 Expression in Lung Cancer

Background

Because most lung cancers are diagnosed at advanced stages, we are forced to conduct molecular testing using small biopsy samples. Endobronchial ultrasound-transbronchial needle aspiration (EBUS-TBNA) is emerging as a minimally invasive biopsy technique. Here, we examined the usefulness of EBUS-TBNA to evaluate programmed death-ligand 1 (PD-L1) expression in lung cancer.

Dissecting the expression of EEF1A1/2 genes in human prostate cancer cells: the potential of EEF1A2 as a hallmark for prostate transformation and progression

Background:

In prostate adenocarcinoma, the dissection of the expression behaviour of the eukaryotic elongation factors (eEF1A1/2) has not yet fully elucidated.

Methods:

The EEF1A1/A2 expressions were investigated by real-time PCR, western blotting (cytoplasmic and cytoskeletal/nuclear-enriched fractions) and immunofluorescence in the androgen-responsive LNCaP and the non-responsive DU-145 and PC-3 cells, displaying a low, moderate and high aggressive phenotype, respectively. Targeted experiments were also conducted in the androgen-responsive 22Rv1, a cell line marking the progression towards androgen-refractory tumour. The non-tumourigenic prostate PZHPV-7 cell line was the control.

Results:

Compared with PZHPV-7, cancer cells showed no major variations in EEF1A1 mRNA; eEF1A1 protein increased only in cytoskeletal/nuclear fraction. On the contrary, a significant rise of EEF1A2 mRNA and protein were found, with the highest levels detected in LNCaP. Eukaryotic elongation factor 1A2 immunostaining confirmed the western blotting results. Pilot evaluation in archive prostate tissues showed the presence of EEF1A2 mRNA in near all neoplastic and perineoplastic but not in normal samples or in benign adenoma; in contrast, EEF1A1 mRNA was everywhere detectable.

Conclusion:

Eukaryotic elongation factor 1A2 switch-on, observed in cultured tumour prostate cells and in human prostate tumour samples, may represent a feature of prostate cancer; in contrast, a minor involvement is assigned to EEF1A1. These observations suggest to consider EEF1A2 as a marker for prostate cell transformation and/or possibly as a hallmark of cancer progression.     

A functional insertion/deletion polymorphism in the promoter region of NFKB1 gene increases susceptibility for nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is a common malignancy in southern China and Southeast Asia. Nuclear factor-κB (NF-κB)-activation plays critical roles in Epstein–Barr virus (EBV) latent membrane protein 1 (LMP1) mediated tumorigenesis in NPC. A functional insertion/deletion polymorphism (−94 insertion/deletion ATTG) in the promoter of NFKB1 gene, which encodes the p50 subunit of NF-κB protein complex, was recently identified. This study found that the frequency of ATTG₂ allele in NPC patients was significantly higher than that in control subjects (66% vs. 57.1%, p = 0.015, OR = 1.453), suggesting that the functional NFKB1 promoter polymorphism is associated with increased risk for NPC.