Juri Sugiyama

Immunosuppression through constitutively activated NF-κB signalling in human ovarian cancer and its reversal by an NF-κB inhibitor

Background:Although T-cell immunity is thought to be involved in the prognosis of epithelial ovarian cancer (EOC) patients, immunosuppressive conditions hamper antitumour immune responses. Thus, their mechanisms and overcoming strategies need to be investigated.Methods:The role of NF-κB in human EOC cells and macrophages was evaluated by in vitro production of immunosuppressive IL-6 and IL-8 by EOC cells and in vivo analysis of immune responses in nude mice implanted with human EOC cells using an NF-κB inhibitor DHMEQ.Results:In EOC patients, increased plasma IL-6, IL-8, and arginase were observed. The NF-κB inhibitor DHMEQ inhibited the production of IL-6 and IL-8 by EOC cell lines. Immunosuppression of human DCs and macrophages by culture supernatant of EOC cells was reversed with the pretreatment of DHMEQ. Administration of DHMEQ to nude mice implanted with human EOC resulted in the restoration of T-cell stimulatory activity of murine DCs along with the reduction of tumour accumulation and arginase expression of MDSCs. Nuclear factor-κB inhibition in tumour-bearing mice also enhanced antitumour effects of transferred murine naive T cells.Conclusions:NF-κB is involved in the immunosuppression induced by human EOC, and its inhibitor may restore antitumour immune responses, indicating that NF-κB is an attractive target for EOC treatment.

Pregnancy Outcomes After Abdominal Radical Trachelectomy for Early-Stage Cervical Cancer: A 13-Year Experience in a Single Tertiary-Care Center.

Objective To investigate pregnancy outcomes in women after abdominal radical trachelectomy (RT) for early-stage cervical cancer. Methods The patients’ background, fertility, and pregnancy outcomes were reviewed in a total of 61 pregnancies in 48 of 172 women who underwent abdominal RT at Keio University Hospital between September 2002 and December 2013. Results There were 5 women with stage IA1, 2 with stage IA2, and 41 with stage IB1. Histological types were as follows: squamous cell carcinoma (n = 36), adenocarcinoma (n = 10), and adenosquamous cell carcinoma (n = 2). The pregnancy rate of women attempting to conceive after abdominal RT was 44% (48/109). The mean ± SD duration from abdominal RT to conception was 3.1 ± 1.9 years. Of 61 pregnancies, 42 pregnancies were achieved by fertility treatment (in vitro fertilization-embryo transfer, 39; intrauterine insemination, 3). After excluding one pregnancy without detailed clinical information, there were 42 live births (5 in 22–27 weeks, 11 in 28–33weeks, 20 in 34–36 weeks, and 6 in 37–38 weeks), 13 miscarriages, and 5 ongoing pregnancies. While there were 10 first trimester miscarriages, 3 pregnancies ended in the second trimester owing to chorioamnionitis. The mean gestational age at birth was 33 weeks of pregnancy. Thirty-seven neonates were appropriate-for-date, and one was small-for-date. Six pregnancies exhibited massive bleeding from the residual cervix in the late pregnancy. Preterm birth less than 34 weeks of pregnancy was related to premature rupture of the membrane (P < 0.05). Chorioamnionitis was evident in 9 of 11 pregnancies with preterm premature rupture of the membrane followed by birth at less than 34 weeks of pregnancy. No parturients exhibited lochiometra and endometritis postpartum. Conclusions Abdominal RT provided favorable pregnancy outcomes, and fertility treatment could be advantageous to conception. Massive bleeding from the residual cervix as well as ascending infection might be characteristic features during pregnancy after abdominal RT.

Aberrant Myosin 1b Expression Promotes Cell Migration and Lymph Node Metastasis of HNSCC.

Lymph node metastasis is the major clinicopathologic feature associated with poor prognosis in patients with head and neck squamous cell carcinoma (HNSCC). Here, web-based bioinformatics meta-analysis was performed to elucidate the molecular mechanism of lymph node metastasis of human HNSCC. Preferential upregulation of Myosin 1b (MYO1B) transcript in HNSCC datasets was identified. Myo1b mRNA was highly expressed in human HNSCC cells and patient tissue specimens compared with their normal counterparts as shown by quantitative PCR (qPCR) analyses. Immunohistochemistry (IHC)-detected Myo1b expression was significantly correlated with lymph node metastases in patients with oral cancer of the tongue. HNSCC with high expression of Myo1b and chemokine receptor 4 (CCR4), another metastasis-associated molecule, was strongly associated with lymph node metastasis. RNA interference (RNAi) of Myo1b in HNSCC cells, SAS and HSC4, significantly inhibited migratory and invasive abilities through decreased large protrusion formation of cell membranes. Finally, Myo1b knockdown in SAS cells significantly inhibited in vivo cervical lymph node metastases in a cervical lymph node metastatic mouse model system. Implications: Myo1b is functionally involved in lymph node metastasis of human HNSCC through enhanced cancer cell motility and is an attractive target for new diagnostic and therapeutic strategies for patients with HNSCC. Mol Cancer Res; 13(4); 721–31. ©2014 AACR.

Repair of the threatened feminine identity: Experience of women with cervical cancer undergoing fertility preservation surgery

Background: Fertility preservation is important for women of reproductive age with cervical cancer. The underlying reasons behind suboptimal reproductive results after successful fertility-preserving surgery have not yet been fully revealed. Objective: The objective of this study was to explore the experience of fertility preservation with radical trachelectomy from the perspective of women with cervical cancer. Methods: We conducted interviews with women with cervical cancer who underwent radical trachelectomy using a Grounded Theory methodology with a theoretical framework of symbolic interactionism. Results: Our findings articulate a process in which feminine identity is first threatened by a diagnosis of cancer, then repaired by fertility preservation with radical trachelectomy, and finally reconstructed after the surgery, through interactions with self, others, and external events in women with cervical cancer. Feeling incomplete as a woman because of the loss of the uterus was a critical factor in the women’s feeling that their feminine identity was threatened. Thus, fertility preservation was significant for these women. The meaning of fertility preservation varied among the women, and their life perspectives were therefore distinct after the surgery. Conclusions: Women with cervical cancer who undergo radical trachelectomy experience an identity transformation process, and child bearing is not the only expected outcome of fertility preservation. Implications for Practice: Nurses should coordinate care through the cancer trajectory. Understanding the identity transformation process helps nurses to assess patients’ needs and provide appropriate individual care.

Enhanced anti-tumor effects of the PD-1/PD-L1 blockade by combining a highly absorptive form of NF-kB/STAT3 inhibitor curcumin

The PD-1/PD-L1 blockade is now recognized as one of the basic immune interventions for development of combination cancer immunotherapy. We have been screening chemical libraries to obtain compounds which have an activity to augment anti-tumor activity of the PD-1/PD-L1 blockade. We have previously shown that activation of NF-kB and STAT3 signals in both cancer cells and immune cells plays important roles in cancer cell-induced immunosuppression, and their inhibition may augment anti-tumor CTL. Curcumin, a major active component of turmeric, has activities to inhibit both NF-kB and STAT3 signals. We have recently developed a highly absorptive form of curcumin which oral administration resulted in high plasma concentration. In this study, we evaluated combination treatment of this highly absorptive form of curcumin with the PD-1/PD-L1 blockade. Curcumin inhibited significantly in vitro production of IL6 and IL8 by human ovarian cancer cell lines (OC) having activated NF-kB and STAT3 signaling. In nude mice implanted with human OC, T-cell stimulatory activity of murine DC was impaired partly through increased human IL-6 from human OC. Systemic p.o. administration of curcumin decreased human IL6 in mouse serum and restored T-cell stimulatory activity of murine DC in spleens and tumors. In two syngeneic murine colon cancer models, NF-kB dependent IL6 producing MC38 in B6 mice and NF-kB non-activated IL6 non-producing CT26 in Balb/C mice, curcumin p.o. administration resulted in the significant augmentation of tumor antigen-specific CD8+ T-cell induction accompanied by increased T-cell stimulatory activity of DC via decreased STAT3 and NF-kB signaling, although curcumin did not inhibit proliferation of both tumor cell lines. Furthermore, p.o. administration of curcumin into non-tumor bearing B6 mice immunized with ovalbumin (OVA) peptide augmented OVA-specific CTL induction. These results indicate that curcumin not only enhances tumor antigen specific T-cell via reversal of tumor-induced immunosuppression (e.g. IL6 induced DC impairment), but also enhances CTL via directly acting on immune cells. Since PD-1 positive T cells were infiltrated in tumors expressing PD-L1, we evaluated combination of curcumin and anti-PD-L1 blockade, and found that combination of curcumin and anti-PD-L1 Ab had synergistic anti-tumor activity. These results indicate that combination of curcumin which enhances induction of tumor antigen specific, PD-1 positive CTL in tumors via acting on both cancer cells and immune cells and local PD-1/PD-L1 blockade, is an attractive strategy for development of effective cancer immunotherapy.

Personalized cancer immunotherapy: Immune biomarkers and combination immunotherapy

Cancer immunotherapies utilizing tumor-specific T-cell responses, immune-checkpoint blockade, and T-cell-based adoptive cell therapy, have recently shown durable responses in advanced patients with various cancers. However, there are still cancer types and patients who do not respond to these immunotherapies. Pretreatment immune status varies in cancer patients, and it correlates with prognosis after various cancer therapies including immunotherapy. The differential T-cell response is defined by positive (e.g., number of immunogenic mutated peptides derived from mainly passenger DNA mutations in cancer cells, polymorphisms of immune-related genes of patients) and negative (e.g., oncogene activation including driver DNA mutations) immune pathways along with environmental factors (e.g., intestinal microbiota, diet, smoking, infection history). These factors could be biomarkers for selection of the patients who are likely to respond to immunotherapy and furthermore could be therapeutic targets to improve efficacy of immunotherapy possibly by combination immunotherapy with interventions on multiple key regulation points in the antitumor T-cell responses. Personalized combination immunotherapy based on the evaluation of T-cell immune status is a promising strategy for cancer treatment.

Development of Personalized Combination Cancer Immunotherapy Based on the Patients’ Immune Status

Cancer immunotherapies, particularly immune-checkpoint blockade and T cell-based adoptive cell therapy, have recently been recognized as cancer treatments that show strong and durable responses even for advanced cancer patients with multiple metastases. The major issues in the development of cancer immunotherapy are the identification of biomarkers to distinguish responders and non-responders, and the improvement of efficacy of immunotherapy possibly by combination with appropriate immune interventions targeting different key regulating points in the anti-tumor immune responses. Interestingly, pretreatment T cell immune status varies among cancer patients, and appears to correlate with responses to various cancer treatments including surgery, chemotherapy, radiation therapy, and immunotherapy. Balance of anti-tumor T cell induction pathway and immunosuppressive pathway, which are regulated by characteristics of both cancer cells and patients’ immune reactivity, may define the differential immune status among cancer patients along with environmental factors such as intestinal microbiota. The analysis of such mechanisms may lead to the identification of immune biomarkers and immune-modulating strategies for combination immunotherapies. Further research on human cancer immunopathology will lead to the development of effective personalized combination immunotherapies based on the evaluation of cancer patients’ immune status.

Cancer Induced Immunosuppression and Its Modulation by Signal Inhibitors

Although cancer immunotherapy has recently demonstrated durable responses even in patients with advanced cancer, not all patients or cancer types respond to the therapy. Pretreatment immune status varies among cancer patients and is correlated with responses to immunotherapy. Immune conditions may be defined by the balance of positive and negative pathways in the anti-tumor immune responses, which are regulated by both cancer cell characteristics and patients’ immune-reactivity along with various environmental factors. Gene alterations and signal activation define the immunological characteristics of cancer cells; tumor specific peptides derived from passenger mutations induce anti-tumor T-cells and oncogene activation (e.g. driver mutations, overexpression: MAPK, STAT3, NF-κB, β-catenin) rather promote immunosuppression. Oncogene/signal activation in cancer cells triggers multiple immunosuppressive cascades involving various immunosuppressive molecules and cells (e.g. TGF-β, IL10, IL6, VEGF, Treg, MDSC). Signal inhibitors are able to augment anti-tumor T-cell responses through multiple mechanisms including inhibition of cancer-induced immunosuppression, immunogenic cancer cell death, and enhancement of immune cell functions. Since the oncogene-signal activation status is different among patients, personalized immunotherapy combined with appropriate signal inhibitors may be considered for the development of effective immunotherapy.

Immunosuppression through HNF-1β signaling in human ovarian clear cell cancer

Cancer-induced immunosuppression is one of the major problems for development of cancer immunotherapies. A transcriptional factor HNF-1β preferentially activated in human ovarian clear cell cancer (OCCC) was reported to contribute to various malignant features including metastases and glucose metabolism. In this study, we have investigated roles of HNF-1β in the immunosuppressive activity of human OCCC. HNF-1β knockdown and overexpression experiments revealed that HNF-1β induced production of IL6 and IL8, which were elevated in OCCC patient plasma. HNF-1β was found to promote production of IL6 and IL8 through activated STAT3 signaling and NF-κB dependent osteopontin pathway. In vitro suppressive activities of human OCCC culture supernatants on generation of human monocyte-derived dendritic cell (DC) were reduced by siRNA knockdown of HNF-1β in cancer cells partly through decrease of IL6 production. In the nude mice implanted with human OCCC cell lines, knockdown of HNF-1β in the cancer cells resulted in restoration of T cell stimulatory activity of murine splenic DC, and decrease of accumulation and arginase expression of myeloid-derived suppressor cells in spleens and tumors accompanied by human IL6 decrease. In the OCCC patient plasma, IL8 levels were correlated with the levels of immunosuppressive arginase, indicating that IL8 may also be involved in immunosuppression. Therefore, HNF-1β activation in human OCCC is an upstream event for induction of immunosuppression via STAT3 and NF-κB activation, and is an attractive target for restoring immunocompetence in OCCC patients.