Serine Protease Inhibitor Kazal Type 1, A Potential Biomarker for the Early Detection, Targeting, and Prediction of Response to Immune Checkpoint Blockade Therapies in Hepatocellular Carcinoma.

Background: We aimed to characterize serine protease inhibitor Kazal type 1 (SPINK1) as a gene signature for the early diagnosis, molecular targeting, and prediction of immune checkpoint blockade (ICB) treatment response of hepatocellular carcinoma (HCC). Methods: The transcriptomics, proteomics, and phenotypic analyses were performed separately or in combination. Results: We obtained the following findings on SPINK1. Firstly, in the transcriptomic training dataset, which included 279 stage I and II tumor samples (out of 1,884 stage I-IV HCC specimens) and 259 normal samples, significantly higher area under curve (AUC) values and increased integrated discrimination improvement (IDI) and net reclassification improvement (NRI) were demonstrated for HCC discrimination in SPINK1-associated models compared with those of alpha-fetoprotein (AFP). The calibration of both SPINK1-related curves fitted significantly better than that of AFP. In the two independent transcriptomic validation datasets, which included 201, 103 stage I-II tumor and 192, 169 paired non-tumor specimens, respectively, the obtained results were consistent with the above-described findings. In the proteomic training dataset, which included 98 stage I and II tumor and 165 normal tissue samples, the analyses also revealed better AUCs and increased IDI and NRI in the aforementioned SPINK1-associated settings. A moderate calibration was shown for both SPINK1-associated models relative to the poor results of AFP. Secondly, in the in vitro and/or in vivo murine models, the wet-lab experiments demonstrated that SPINK1 promoted the proliferation, clonal formation, migration, chemoresistance, anti-apoptosis, tumorigenesis, and metastasis of HCC cells, while the anti-SPINK1 antibody inhibited the growth of the cells, suggesting that SPINK1 has "tumor marker" and "targetable" characteristics in the management of HCC. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that SPINK1 was engaged in immunity-related pathways, including T-cell activation. Thirdly, in the transcriptomic analyses of the 368 HCC specimens from The Cancer Genome Atlas (TCGA) cohort, the high abundance of SPINK1 was positively correlated with the high levels of activated tumor-infiltrating CD4+ and CD8+ T lymphocytes and dendritic and natural killer cells, while there were also positive correlations between SPINK1 and immune checkpoints, including PD-1, LAG-3, TIM-3, TIGIT, HAVCR2, and CTLA-4. The ESTIMATE algorithm calculated positive correlations between SPINK1 and the immune and ESTIMATE scores, suggesting a close correlation between SPINK1 and the immunogenic microenvironment within HCC tissues, which may possibly help in predicting the response of patients to ICB therapy. Conclusions: SPINK1 could be a potential biomarker for the early detection, targeted therapy, and prediction of ICB treatment response in the management of HCC.

Enhancement of gemcitabine efficacy by K73-03 via epigenetically regulation of miR-421/SPINK1 in gemcitabine resistant pancreatic cancer cells.

BACKGROUND: Gemcitabine (GCB) is a first-line chemotherapeutic drug for pancreatic cancer (PCa). However, the resistance begins developing within weeks of chemotherapy. SPINK1 overexpression enhances resistance to chemotherapy. In a recent study, our laboratory established that the oleanolic acid (OA) derivative, K73-03, had a strong inhibitory effect on a SPINK1 overexpressed PCa cells. PURPOSE: In our current study, we studied the enhancement of GCB inhibitory effect by K73-03, a new novel OA derivative, alone or in combination with GCB on the GCB-resistant PCa cells by mitochondrial damage through regulation of the miR-421/SPINK1. METHODS: We detected the binding between miR-421 and SPINK1-3'-UTR in GCB-resistant PCa cells using Luciferase reporter assays. Cells viability, apoptosis, migration, and mitochondrial damage were investigated. RESULTS: The results demonstrated that the combination of K73-03 and GCB suppressed the growth of AsPC-1 and MIA PaCa-2 cells synergistically, with or without GCB resistance. Mechanistic findings showed that a combination of K73-03 and GCB silences SPINK1 epigenetically by miR-421 up-regulating, which leads to mitochondrial damage and inducing apoptosis in GCB-resistant PCa cells. CONCLUSION: We found an interesting finding that the 73-03 in combination with GCB can improve GCB efficacy and decrease PCa resistance, which induced apoptosis and mitochondrial damage through epigenetic inhibition of SPINK1 transcription by miR-421 up-regulation. This was the first study that used OA derivatives on GCB-resistant PCa cells, so this combined strategy warrants further investigation.

Outcomes of local thoracic surgery in patients with stage IV non-small-cell lung cancer: A SEER-based analysis.

BACKGROUND: The outcomes of thoracic surgery for patients with stage IV non-small-cell lung cancer (NSCLC) are controversial and uncertain. PATIENTS AND METHODS: The National Cancer Institute's Surveillance, Epidemiology, and End Results was queried for patients with stage IV NSCLC, including those treated with surgery-participated therapy modalities. Overall survival (OS) was evaluated using a variety of statistical analyses. RESULTS: The analysis was carried out for 90,982 patients from 1975 to 2016 who had been diagnosed as stage IV NSCLC. Propensity score-matched (PSM) analyses that were well-balanced with all the important confounding covariates revealed improved OS (median survival time [MST]) with patients receiving surgery versus non-surgery (MST: 15 versus 8 months, P < 0.001); undergoing surgery plus chemotherapy versus chemotherapy (MST: 19 versus 11 months, P < 0.001); and having surgery plus chemoradiation versus chemoradiation (MST: 18 versus 11 months, P < 0.001). Sequential landmark analyses for long-term survivors of ≥1 and ≥3 years all indicated improved OS (P < 0.001) on univariate and multivariate analyses for the patients receiving the three surgery-related treatment patterns listed earlier, relative to the corresponding surgery-absent treatment modalities. For synchronous presentations of varied treatment paradigms, surgical intervention significantly led to increased OS (MST, months) benefits following treatment paradigms: surgery plus chemotherapy (22), surgery plus chemoradiation (18), chemotherapy (10), surgery only (9), chemoradiation (9), surgery plus radiation (6) and radiation alone (2). The subgroup analysis demonstrated that the elevated OS associated with local thoracic surgery in addition to chemotherapy (versus chemotherapy) or chemoradiation (versus chemoradiation) fell in the subcategories of T0-3, N0-2 and 0-1 (metastatic sites) tumours. The comparison of the aforementioned two types of treatment patterns indicated that the optimal patients for the surgery were those with any combination of T1-4, N0-3, Msite0-1 and adeno- or squamous carcinoma. CONCLUSIONS: The patients with T1-4, N0-3, Msite0-1 and adeno- or squamous carcinoma of stage IV NSCLC had a longer OS with local thoracic surgery in combination with chemotherapy or chemoradiation.