Efficacy of the induced pluripotent stem cell derived and engineered CD276-targeted CAR-NK cells against human esophageal squamous cell carcinoma.

Introduction: Chimeric antigen receptor natural killer (CAR-NK) cells have been found to be successful in treating hematologic malignancies and present potential for usage in solid tumors. Methods: In this study, we created CD276-targeted CAR-expressing NK cells from pluripotent stem cells (iPSC CD276-targeted CAR-NK cells) and evaluated their cytotoxicity against esophageal squamous cell carcinoma (ESCC) using patient-specific organoid (PSO) models comprising of both CD276-positive and CD276-negative adjacent epithelium PSO models (normal control PSO, NC PSO) as well as primary culture of ESCC cell models. In addition, in vitro and in vivo models such as KYSE-150 were also examined. iPSC NK cells and NK-free media were used as the CAR-free and NK-free controls, respectively. Results: The positive CD276 staining was specifically detected on the ESCC membrane in 51.43% (54/105) of the patients of all stages, and in 51.35% (38/74) of stages III and IV. The iPS CD276-targeted CAR-NK cells, comparing with the iPS NK cells and the NK-free medium, exhibited specific and significant cytotoxic activity against CD276-positive ESCC PSO rather than CD276-negative NC PSO, and exhibited significant cytotoxicity against CD276-expressing cultured ESCC cells, as well as against CD276-expressing KYSE-150 in vitro and in BNDG mouse xenograft. Discussion: The efficacy of the iPSC CD276-targeted CAR-NK cells demonstrated by their successful treatment of CD276-expressing ESCC in a multitude of pre-clinical models implied that they hold tremendous therapeutic potential for treating patients with CD276-expressing ESCC.

Efficacy of MUC1-targeted CAR-NK cells against human tongue squamous cell carcinoma.

Introduction: The clinical efficacy of CAR-NK cells against CD19-expressing blood cancers has been demonstrated, and they have shown potential for treating solid tumors as well. However, the efficacy of CAR-NK cells for treating human oral tongue squamous cell carcinoma (OTSCC) has not been examined. Methods: We assessed MUC1 expression in human OTSCC tissue and a cell line using immunohistochemistry and immunofluorescence. We constructed NK cells that express CAR targeted to MUC1 from pluripotent stem cells (iPSC-derived MUC1-targeted CAR-NK cells) and evaluated their effectiveness against OTSCC in vitro using the xCELLigence Real-Time Cell Analysis system and CCK8 assay, and in vivo by measuring xenograft growth daily in BNDG mice treated with MUC1-targeted CAR-NK cells. As controls, we used iPSC-derived NK cells and NK-free media, which were CAR-free and blank, respectively. Results: MUC1 expression was detected in 79.5% (66/83) of all OTSCC patients and 72.7% (24/33) of stage III and IV. In stage III and IV MUC1 positive OTSCC, 63.6% (21/33) and 48.5% (16/33) patients had a MUC1-positive cancer cell rate of more than 50% and 80%, respectively. The iPSC-derived MUC1-targeted CAR-NK cells exhibited significant cytotoxicity against MUC1-expressing OTSCC cells in vitro, in a time- and dose-dependent manner, and showed a significant inhibitory effect on xenograft growth compared to both the iPSC-derived NK cells and the blank controls. We observed no weight loss, severe hematological toxicity or NK cell-mediated death in the BNDG mice. Conclusion: The MUC1-targeted CAR-NK cells had significant efficacy against human OTSCC, and their promising therapeutic response warrants further clinical trials.

Identification and validation of a novel necroptosis-related molecular signature to evaluate prognosis and immune features in breast cancer.

Necroptosis has been shown to play an important role in the development of tumors. However, the characteristics of the necroptosis-related subtypes and the associated immune cell infiltration in the tumor microenvironment (TME) of breast cancer (BRCA) remain unclear. In this study, we identified three clusters related to necroptosis using the expression patterns of necroptosis-relevant genes (NRGs), and found that these three clusters had different clinicopathological features, prognosis and immune cell infiltration in the TME. Cluster 2 was characterized by less infiltration of immune cells in the TME and was associated with a worse prognosis. Then, a necroptosis risk score (NRS) composed of 14 NRGs was constructed using the least absolute shrinkage and selection operator regression (LASSO) Cox regression method. Based on NRS, all BRCA patients in the TCGA datasets were classified into a low-risk group and a high-risk group. Patients in the low-risk group were characterized by longer overall survival (OS), lower mutation burden, and higher infiltration level of immune cells in the TME. Moreover, the NRS was significantly associated with chemotherapeutic drug sensitivity. Finally, the knockdown of VDAC1 reduced the proliferation and migration of BRCA cells, and promoted cell death induced by necroptosis inducer. This study identified a novel necroptosis-related subtype of BRCA, and a comprehensive analysis of NRGs in BRCA revealed its potential roles in prognosis, clinicopathological features, TME, chemotherapy, tumor proliferation, and tumor necroptosis. These results may improve our understanding of NRGs in BRCA and provide a reference for developing individualized therapeutic strategies.

BME-free primary patient-specific organoids obtained with a one-day mimicking method to replicate the corresponding tumor for personalized treatment options.

Introduction: In cancer treatment, every minute counts. Due to the unpredictable behavior of cancer cells caused by continuous mutations, each cancer patient has a unique situation and may or may not respond to a specific drug or treatment. The process of finding an effective therapy can be time-consuming, but cancer patients do not have the luxury of time for trial and error. Therefore, a novel technology to fast generate a patient relevant organoid for the therapies selecting is urgently needed. Methods: Utilizing the new organoid technology by specially dissolving the mesenchyme in tumor tissues acquired from cancer patients, we realized the work of creating patient-specific organoids (PSO) within one day. Results: PSO properties reflect those of its respective original in vivo tumor tissue and can be utilized to perform various in vitro drug sensitivity tests to identify the most effective clinical treatment for patients. Additionally, PSO can aid in assessing the efficacy of immune cell therapies. Discussion: Organoid technology has advanced significantly in recent years. However, current cancer organoid methods involve creating 3D tumor tissue from 2D cancer cells or cell clusters, primarily for cancer research purposes aimed at investigating related molecular and cellular mechanisms of tumor development. These methods are research-driven, not tailored towards clinical applications, and cannot provide personalized information for individual patients. PSO filled the gap of clinic-driven and time-saving method for the personalized therapies selecting to the cancer patients.

Postoperative Systemic Immune-Inflammation Index (SII): A Superior Prognostic Factor of Endometrial Cancer.

Objective: This study evaluates the preoperative and postoperative systemic immune-inflammation index (SII) capacity to predict the prognosis of patients with endometrial carcinoma after the operation and build a nomogram model to assist clinical practice. Methods: The retrospective study included 362 consecutive patients with surgically resected endometrial cancer between January 2010 and June 2015 at The Affiliated Cancer Hospital of Shantou University Medical College. Blood routine was examined within 1 week before surgery to calculate SII, NLR, PLR, and MLR and 3 days after surgery to measure SII. The Pearson's χ2-test or Fisher's exact test was used to explore their relationship to clinical variables. The univariate and multivariate survival analyses were performed by Cox regression to identify the independent prognostic indicators. The Kaplan-Meier method with the log-rank test was used to generate the overall survival (OS) curves. R software was used to generate the receiver operating characteristic (ROC) curve and then it got the optimum cutoff value through the maximum Youden index. A nomogram model was formed with systemic immune inflammation and clinical factors. Results: The preoperative SII was related to age (p = 0.009), FIGO stage (p = 0.02) and menopause (p = 0.014). The postoperative SII was associated with menopause (p = 0.014). Univariate analysis indicated that FIGO stage, lymphatic invasion, depth of myometrial invasion, postoperative chemotherapy, postoperative radiotherapy, preoperative SII, NLR, PLR, MLR, CRP, CA125, and postoperative SII were predictors of OS (p < 0.05). Multivariate analysis showed that lymphatic invasion and postoperative SII were independent prognostic factors of OS (p < 0.05). The nomogram model was visualized precisely to reflect the prognosis with a C-index value of 0.866 in this model. Conclusion: The postoperative SII is the independent prognostic factor in patients with endometrial carcinoma after the operation and contributes to poor outcomes. However, after surgery, the preoperative SII and preoperative NLR, PLR, and MLR are not associated with OS endometrial carcinoma. Making good use of the nomogram model would contribute to better subsequent therapy.

The Nuclear Farnesoid X Receptor Reduces p53 Ubiquitination and Inhibits Cervical Cancer Cell Proliferation.

The role of farnesoid X receptor (FXR) in cervical cancer and the underlying molecular mechanism remain largely unknown. Therefore, this study aimed to assess the mechanism of FXR in cervical cancer. Western blot, qRT-PCR, and immunohistochemistry demonstrated that FXR was significantly reduced in squamous cell carcinoma tissues, although there were no associations of metastasis and TNM stage with FXR. In Lenti-FXR cells obtained by lentiviral transfection, the overexpression of FXR reduced cell viability and colony formation. Compared with the Lenti-Vector groups, the overexpression of FXR induced early and late apoptosis and promoted G1 arrest. With time, early apoptosis decreased, and late apoptosis increased. In tumor xenograft experiments, overexpression of FXR upregulated small heterodimer partner (SHP), murine double minute-2 (MDM2), and p53 in the nucleus. Co-immunoprecipitation (Co-IP) showed that SHP directly interacted with MDM2, which is important to protect p53 from ubiquitination. Nutlin3a increased MDM2 and p53 amounts in the Lenti-Vector groups, without effects in the Lenti-FXR groups. Silencing SHP reduced MDM2 and p53 levels in the Lenti-FXR groups, and Nutlin3a counteracted these effects. Taken together, these findings suggest that FXR inhibits cervical cancer via upregulation of SHP, MDM2, and p53.

The variable association between expression and methylation of estrogen receptors and the survival of patients with different tumors.

BACKGROUND: Estrogen receptor (ER) is essential in reproductive development and is also the primary driver of breast cancers. Deregulation of ER may also be involved in tumorigenesis of other organs. To understand the role of ER in different tumor types, pan-cancer analysis of estrogen receptor alpha (ESR1) and estrogen receptor beta (ESR2) in various tumors and association with patients' survival were conducted using The Cancer Genome Atlas (TCGA) data. RESULTS: Gene methylation level was evaluated by the mean methylation level of CpG sites in the promoter region. The significant different DNA methylation between tumor and healthy tissues was shown in 10 tumor types for ESR1 and eight tumor types for ESR2. The methylation pattern was also varied across different TCGA tumors. The pan-cancer analysis showed significantly different mRNA expression of ESR1 in nine tumor types and ESR2 in four tumor types. Survival analysis showed that the effects of ERs expression on survival are diverse in different tumors. The expression of ERs was associated with tumor molecular subtypes and various clinical characteristics. ER correlated genes were mainly enriched in cancer and immune-related pathways. CONCLUSIONS: Our pan-cancer analysis data indicated that ERs might be significantly associated with carcinogenesis and progression of some tumors, which may be potential therapeutic targets and prognosis biomarkers.

[Clinicopathological characteristics and prognosis of uterine malignant mixed Mullerian tumor].

OBJECTIVE: To evaluate the clinicopathological characteristics, treatment and prognosis of uterine malignant mixed mullerian tumor. METHODS: The clinical, pathologic and follow-up data of 16 patients with uterine malignant mixed Mullerian tumor treated in our hospital between March, 2003 and June, 2015 were analyzed. RESULTS: The 16 patients had a median age of 58 years at diagnosis, and 13 of them were postmenopausal. The number of patients with FIGO stage Ia, Ib, II, IIIa, IIIc2, and IV was 7, 3, 1, 3, 1, and 1, respectively. In 15 patients who received uterine segment diagnostic curettage, pathological examination all reported malignant results. Among the 15 patients having serum CA125 level test upon admission, 2 had elevated CA125 levels. The overall and disease-free survival rates of the 16 patients were 75% and 68.8%, respectively, and the 3-year survival rate of 13 patients who were followed up for at least 3 years was 72.7%. Two out of 12 patients receiving retroperitoneal lymph?node?dissection?had had postoperative recurrence, as compared with 3 out 4 who did not had the operation; tumor recurrence was found in 3 out of 13 patients receiving postoperative chemotherapy, as compared with 2 out of 3 patients who did not have chemotherapy; tumor recurrence occurred in 1 out of 10 patients receiving radiotherapy, as compared with 4 out of 6 patients without radiotherapy. The recurrence rates in 11 patients with FIGO stage I-II was 18.2%, and that among the 5 patients with FIGO stage III-IV was 60.0%. CONCLUSIONS: Uterine segment diagnostic curettage has a high diagnostic value for uterine malignant mixed Mullerian tumor. FIGO stage is the important prognostic factor for these patients, and early?diagnosis, accurate surgical staging, platinum-based chemotherapy and postoperative pelvic radiotherapy are all associated with a better prognosis.

Identification of a novel isoform of DHRS4 protein with a nuclear localization signal.

The DHRS4 gene encodes an NADP(H)-dependent retinol dehydrogenase/reductase (NRDR) and plays an important role in regulating the synthesis of retinoic acid. In the present study, we identified a novel splice RNA variant, designated NRDRA2, of the human DHRS4 gene by RT-PCR, 3' RACE, and 5' RACE. NRDRA2 mRNA lacked exons 4 and 6, and had a shift in the reading frame when compared to DHRS4 mRNA, resulting in loss of the peroxisomal targeting signal of NRDR and gain of a nuclear localization signal in the predicted NRDRA2 protein. Endogenous NRDRA2 protein was identified in the human cervical carcinoma cell line HeLa by immunoprecipitation and mass spectrometric assay. A green fluorescent protein reporter assay showed that NRDRA2 protein mainly localized to the nuclei, confirming the sequence at its C-terminus as a legitimate nuclear localization signal sequence. This study identifies the alternative transcript variant NRDRA2 encoding a subcellular nuclear localized NRDRA2 protein.