Critical Reviews in Immunology

Single-Cell Sequencing Combined with RNA Sequencing Reveals the Role of Natural Killer Cells in Prognosis and Immunotherapy Response in Cervical Squamous Cell Carcinoma

Cervical squamous cell carcinoma (CSCC) has an unfavorable prognosis with major therapeutic challenges. Natural killer (NK) cells play a pivotal function in anti-tumor immunity. However, the correlation between NK cells and heterogeneity and prognosis in CSCC lacks definitive understanding. This study seeks to elucidate the potential value of high-activity NK cell-related genes in prognosis and immunotherapy for CSCC. Transcriptome and single-cell sequencing data of people with CSCC were obtained from TCGA and EMBL-EBI databases, respectively. Single-cell data underwent quality control, dimensionality reduction, and identification of high-activity NK cells and their marker genes. After WGCNA application to screen NK-related genes. a prognostic risk model was constructed employing univariate Cox, LASSO Cox regression, and multivariate Cox regression analyses. The clinical implication of the model was validated through immune infiltration assessment, survival, gene set enrichment, tumor mutation analyses, and drug sensitivity prediction. High-activity NK cells and associated genes in CSCC were identified. A risk prognostic model based on high-activity NK-related genes was developed, yielding six key prognostic genes (RIPOR2, PTGER4, BIN2, MARCHF2, SPATA13, KLRC2). The model demonstrated robust predictive performance in training and validation sets. Patients in the low-risk group exhibited higher infiltration levels of NK, CD8<sup>+</sup> T, and dendritic cells, along with increased sensitivity to immune checkpoint inhibitor therapy. Additionally, drug sensitivity analysis identified promising therapeutic candidates. This study, integrating single-cell and RNA sequencing, revealed the heterogeneity of NK cells in CSCC. The risk prognostic model provided prognostic biomarkers and therapeutic targets for CSCC patients, offering a theoretical foundation for immunotherapy research.

STROBE-Cross-Population Mendelian Randomization Analysis of Circulating Inflammatory Proteins and Cervical Cancer Risk in European and Asian Populations

<b>Objective:</b> This study investigates the causal relationship between circulating inflammatory proteins and cervical cancer risk in European and Asian populations using Mendelian randomization (MR), providing insights into inflammation's role in cervical cancer pathogenesis. <b>Method:</b> Data from 91 circulating inflammatory proteins from 11 cohorts (14,824 European participants, 909 cervical cancer cases, 238,249 controls; 605 cases, 89,731 controls in the Asian cohort) were analyzed. Inverse variance weighted (IVW) and MR-Egger methods were used to explore causal relationships. Sensitivity analyses, including Cochran's Q tests and leave-one-out analysis, ensured result reliability. <b>Results:</b> In the European population, higher levels of CCL19, IL-1α, and IL-12B were associated with increased cervical cancer risk, while LIFR and PD-L1 were protective. In the Asian population, elevated CCL19, IL-1α, SLAM, and IL-10Rβ increased risk, while CXCL11, SULT1A1, and CXCL1 showed protective effects. Sensitivity analyses confirmed the robustness of these findings. <b>Conclusion: </b>This study demonstrates a causal relationship between circulating inflammatory proteins and cervical cancer risk in both European and Asian populations. The findings highlight both pro-cancer and protective roles of specific inflammatory proteins, offering insights for biomarkers in cervical cancer risk assessment and prevention strategies.

Cellular Immune Response to High-Risk Human Papillomavirus Infection: A Systematic Review

The relationship between human papillomavirus (HPV) and immune cells is vital for understanding the pathophysiology of infection and its role in neoplastic progression. High-risk human papillomavirus (HR-HPV) is the main cause of cervical cancer (CC). Thus, the association between immune response cells, the virus, and its behavior according to cervical disease development could provide new ways for understanding the entire process. Since the role and the presence of the immune response cells in the uterus cervix considering HPV infection has not been elucidated so far, this study aimed to identify the immune cells involved in high-grade intraepithelial lesions (HSIL) and CC development related to uterine cervical infection caused by HR-HPV. The study population included women who had positive molecular tests for HPV. Through the databases MEDLINE, EMBASE, LILACS, Cochrane, Scopus, Web of Science, CINAHL, Science Direct, and Google Scholar we identified 6,698 studies at the beginning. After the systematic review steps, the final number of included studies was 22. Cervical lesions were distributed according to the severity of lesions in HSIL, low-grade squamous intraepithelial lesions (LSIL), and negative for intraepithelial lesions or malignancy (NILM). The cellular phenotypes presented in these publications were T lymphocytes (LT), regulatory T lymphocytes (Tregs), macrophages (MØ), natural killer cells (NK), natural killer T cells (NKT), Langerhans cells (LC), and dendritic cells (DC). Among the observed associations with cervical lesions and HR-HPV, we highlight the DC/LC and MØ being 36.4% of the cell types, followed by Tregs (31.8%) and LT CD4 / CD8 with 27.3%. The increased findings in innate and adaptive immunological response may imply both are acting together, with the innate response cells and Tregs being the most prominent. Since these cells have great importance in the maintenance and balance of the immunological system, the present study highlights the essential role of MØ and Treg cells in the process of cervical lesion severity associated with HPV, suggesting that they may be focused as prognostic markers and immunotherapeutic targets.

Dual Inhibition of IRE1α and YAP Signaling as a Potential Therapy for Epithelial Ovarian Carcinoma

Epithelial ovarian carcinoma (EOC) is a highly lethal gynecological malignancy with limited treatment options. This study aimed to explore the regulatory roles of IRE1α and YAP1 in EOC progression and identify potential therapeutic targets. Blood and tissue samples were collected from 26 EOC patients and 10 patients with ovarian cysts. The expression of inflammatory factors in the blood was measured using ELISA. The proliferation, migration, invasion, and cell cycle of human ovarian cancer cell lines SK-OV-3, SW626, and Anglene were evaluated using MTT assays, scratch tests, Transwell assays, and flow cytometry. The effects of IRE1α inhibition on EOC cell proliferation, migration, and apoptosis were investigated using pharmacological inhibitors and shRNA knockdown. IRE1α was highly expressed in EOC patients and was negatively correlated with patient survival rates. Additionally, IRE1α scores in EOC patients were positively correlated with serum levels of TNF-α and VEGF-a. Compared to normal controls, significantly higher expressions of IRE1α and XBP1 were observed in ovarian cancer tissues and cells. Knockdown of IRE1α in ovarian cancer cells led to a significant reduction in the expression of IRE1α and XBP1s, as well as inhibited cell proliferation and survival. The IRE1α inhibitors STF-083100 and 4μ8C suppressed the proliferation, invasion, and migration of SK-OV-3 cells and reduced the expression levels of related factors. 4μ8C inhibited the degradation of YAP within SK-OV-3 cells while downregulating the expression of Cyclin D1 protein. Compared to the group treated with 4μ8C alone, the combined intervention of 4μ8C and a YAP inhibitor showed a more pronounced inhibitory effect on the proliferation of SK-OV-3 cells.This study first reveals that the IRE1α/YAP signal drives the malignant progression of EOC through the regulation of cell proliferation, migration, and invasion. The dual-targeted synergistic inhibition of IRE1α/YAP1 offers an innovative therapeutic paradigm for the treatment of EOC.

Commentary: Ovarian Cancer: Path to Effective Treatments

Despite advancements in cancer therapeutics such as checkpoint inhibitors and some targeted therapies, we have not achieved success in effectively treating ovarian cancer, since these therapeutics only benefit a subset of patients, and also provide short-term protection. The use of chemotherapy and radiation therapy can cause depletion and/or lack of immune cells' function. Chimeric antigen receptor T (CAR-T) cell therapy is found to be effective against several blood-based cancers, but limited success was seen against solid tumors. Targeting fewer antigens and significant side effects of therapy decreases the efficacy of CAR-T cells as immunotherapeutic in solid tumors, even though there is a great drive and significant effort to establish these therapies around the world. Bispecific and tri-specific antibodies have recently been advocated as effective cancer therapeutics. However, at present, these also suffer the fate of CAR-Ts since the loss of antigen on tumor cells will render these therapeutics ineffective. At present, we should design therapeutics that may have synergistic effects on killing/treating tumors. The only way we can establish that will be by learning the mechanisms of actions of immune therapeutics. Thus, advancement in the knowledge and effective strategies are required to develop cancer immuno-therapeutics. We have dedicated our efforts to understand the immunobiology of natural killer (NK) cells. One of our most important discoveries was demonstration of targeting of cancer stem-like cells (CSCs)/poorly differentiated tumors exhibiting lower major histocompability complex class I expression by the NK cells. In addition, we showed that supercharged NK (sNK) cells had great ability to target both CSCs/poorly differentiated and well differentiated ovarian tumors, whereas activated primary NK cells only targeted CSCs/poorly differentiated tumors. Therefore, the use of sNK cells in immunotherapy should result in effective elimination of heterogeneous populations of ovarian tumors.

ZC3H13 Enhances the Malignancy of Cervical Cancer by Regulating m6A Modification of CKAP2

Sustained expression of zinc finger CCCH-type containing 13 (ZC3H13) in tumors is essential for cancer cell malignancy; however, our understanding of its clinical effects and mechanisms in cervical cancer (CC) is limited. In this study, we aimed to reveal the effect on CC progression of ZC3H13-mediated N6-methyladenosine (m6A) modification to stabilize cytoskeleton-associated protein 2 (CKAP2) expression. CC tissues and paired adjacent normal tissues were collected from 50 patients. qRT-PCR was used to clarify ZC3H13 and CKAP2 expression levels in the CC tissues. The functional roles of ZC3H13 and CKAP2 in CC were analyzed by detecting the changes in CC cell proliferation, migration, invasion, and tumor growth <i>in vivo</i>. The regulatory relationship between ZC3H13 and CKAP2 was investigated by confirming m6A modification levels and their expression correlation. ZC3H13 and CKAP2 were highly expressed in CC and linked with poor prognosis. We observed that ZC3H13 inhibition decreased CC cell proliferation, invasion, and migration, while its facilitation promoted CC cell malignancy. ZC3H13 mediated m6A modification of CKAP2 to enhance CKAP2 expression in CC cells. Furthermore, CKAP2 overexpression partially restored the malignant phenotypic promotion induced by ZC3H13 overexpression in CC cells. In summary, this study revealed that ZC3H13-mediating m6A modification of CKAP2 promotes CC development. This finding should be conducive to an understanding of the role of ZC3H13-m6A-CKAP2 in CC and should provide an effective therapeutic target for this cancer.

Supercharged NK Cell-Based Immuotherapy in Humanized Bone Marrow Liver and Thymus (Hu-BLT) Mice Model of Oral, Pancreatic, Glioblastoma, Hepatic, Melanoma and Ovarian Cancers

In this paper, we review a number of <i>in vitro</i> and <i>in vivo</i> studies regarding the efficacy of supercharged NK (sNK) cell therapy in elimination or treatment of cancer. We have performed studies using six different types of cancer models of oral, pancreatic, glioblastoma, melanoma, hepatic and ovarian cancers using hu-BLT mice. Our <i>in vitro</i> studies demonstrated that primary NK cells preferentially target cancer stem-like cells (CSCs)/poorly differentiated tumors whereas sNK cells target both CSCs/poorly-differentiated and well-differentiated tumors significantly higher than primary activated NK cells. Our <i>in vivo</i> studies in humanized-BLT mice showed that sNK cells alone or in combination with other cancer therapeutics prevented tumor growth and metastasis. In addition, sNK cells were able to increase IFN-<i>γ</i> secretion and cytotoxic function by the immune cells in bone marrow, spleen, gingiva, pancreas and peripheral blood. Furthermore, sNK cells were able to increase the expansion and function of CD8+ T cells both in <i>in vitro</i> and <i>in vivo </i>studies. Overall, our studies demonstrated that sNK cells alone or in combination with other cancer therapeutics were not only effective against eliminating aggressive cancers, but were also able to increase the expansion and function of CD8+ T cells to further target cancer cells, providing a successful approach to eradicate and cure cancer.