Epigenetic remodeling to improve the efficacy of immunotherapy in human glioblastoma: pre-clinical evidence for development of new immunotherapy approaches.

BACKGROUND: Glioblastoma multiforme (GBM) is a highly aggressive primary brain tumor, that is refractory to standard treatment and to immunotherapy with immune-checkpoint inhibitors (ICI). Noteworthy, melanoma brain metastases (MM-BM), that share the same niche as GBM, frequently respond to current ICI therapies. Epigenetic modifications regulate GBM cellular proliferation, invasion, and prognosis and may negatively regulate the cross-talk between malignant cells and immune cells in the tumor milieu, likely contributing to limit the efficacy of ICI therapy of GBM. Thus, manipulating the tumor epigenome can be considered a therapeutic opportunity in GBM. METHODS: Microarray transcriptional and methylation profiles, followed by gene set enrichment and IPA analyses, were performed to study the differences in the constitutive expression profiles of GBM vs MM-BM cells, compared to the extracranial MM cells and to investigate the modulatory effects of the DNA hypomethylating agent (DHA) guadecitabine among the different tumor cells. The prognostic relevance of DHA-modulated genes was tested by Cox analysis in a TCGA GBM patients' cohort. RESULTS: The most striking differences between GBM and MM-BM cells were found to be the enrichment of biological processes associated with tumor growth, invasion, and extravasation with the inhibition of MHC class II antigen processing/presentation in GBM cells. Treatment with guadecitabine reduced these biological differences, shaping GBM cells towards a more immunogenic phenotype. Indeed, in GBM cells, promoter hypomethylation by guadecitabine led to the up-regulation of genes mainly associated with activation, proliferation, and migration of T and B cells and with MHC class II antigen processing/presentation. Among DHA-modulated genes in GBM, 7.6% showed a significant prognostic relevance. Moreover, a large set of immune-related upstream-regulators (URs) were commonly modulated by DHA in GBM, MM-BM, and MM cells: DHA-activated URs enriched for biological processes mainly involved in the regulation of cytokines and chemokines production, inflammatory response, and in Type I/II/III IFN-mediated signaling; conversely, DHA-inhibited URs were involved in metabolic and proliferative pathways. CONCLUSIONS: Epigenetic remodeling by guadecitabine represents a promising strategy to increase the efficacy of cancer immunotherapy of GBM, supporting the rationale to develop new epigenetic-based immunotherapeutic approaches for the treatment of this still highly deadly disease.

The COVID-19 Pandemic Impact on the Outcome of Medically Assisted Reproduction Pregnancies.

Background: The impact of the Coronavirus Disease-2019 (COVID-19) pandemic on pregnancy is not well-understood. During the outbreak, the initial approach suggested by the major societies was to postpone all non-urgent assisted reproductive technology (ART) treatments. Nevertheless, the Italian Society of Fertility and Sterility and Reproductive Medicine considered ethically correct to proceed with ART treatments, as the infertility rate is increasing over time, with a consistent decline in the live birth rate. The objective of our study was to assess the impact of the COVID-19 pandemic on the outcomes of ART pregnancies, in terms of early pregnancy loss, overall success rate, and live birth rate. Methods: We conducted a single-center retro-prospective cohort study. Patients who underwent ART treatments from 1 March 2020 to 28 February 2021 (pandemic ART cohort, pART; n = 749) and from 1 March 2019 to 29 February 2020 (control cohort, CTR; n = 844) were enrolled. The study had a duration of 24 months. Patients underwent baseline severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) nasopharyngeal swab; quantitative serum beta human chorionic gonadotropin (ß-hCG) to assess pregnancy; pelvic transvaginal ultrasound; and follow-up until delivery. The study took place at the ART Center of the University Hospital in Florence, Italy. Results: There were not statistically significant differences on implantation rate (pART 0.348 ± 0.034 vs. CTR 0.365 ± 0.033, CI = 95%, p = 0.49), clinical pregnancy rate (pART 0.847 ± 0.044 vs. CTR 0.864 ± 0.038, CI = 95%, p = 0.56), and ectopic pregnancy rate (pART 0.008 ± 0.011 vs. CTR 0.01 ± 0.011, CI = 95%, p = 0.79). Neither first trimester miscarriage rate was different between the groups (pART 0.224 ± 0.056 vs. CTR 0.213 ± 0.05, CI = 95%, p = 0.77) nor second trimester miscarriage rate (pART 0.018 ± 0.018 vs. CTR 0.019 ± 0.017, CI = 95%, p = 0.95). Live birth rate remained unchanged during the pandemic (pART 0.22 ± 0.03 vs. CTR 0.239 ± 0.029, CI = 95%, p = 0.37) and stable even when compared to our center rate between 2015 and 2019 (pART 0.222 ± 0.03 vs. general rate 0.224 ± 0.014, CI = 95%, p = 0.83). Conclusion: The COVID-19 pandemic did not cause a statistically significant change in the live birth rate and in the pregnancy loss rate. ART during the COVID-19 pandemic can be considered fair and safe, ethically and medically appropriate. Patients and physicians should be reassured that ART pregnancy outcomes do not seem to be jeopardized by the pandemic state.

COVID-19 Vaccination Does Not Affect Reproductive Health Parameters in Men.

With the implementation of COVID-19 vaccine up-take, doubts regarding the impact of immunization on future fertility have begun to emerge. We have examined vaccine safety on male reproductive health. We set up a multicentre (three infertility centers), retrospective study in order to assess semen parameters and fertilization rate of one hundred-six men in a pairwise comparison between the first and second assisted reproduction technology (ART) attempt, performed respectively before and after COVID-19 vaccination. Median time (range) between the first vaccine dose and the second ART cycle was 75 days (39-112). Semen parameters did not change before and after the exposure. Fertilization rate was also similar before and after vaccination. Twenty-five patients (24%) were oligozoospermic before the vaccination while 26 (25%) after the exposure (P = 0.87). Severe asthenozoospermia were present in 11 patients before as well as after the exposure. No difference was observed even after considering different types of vaccines (mRNA or viral vector). COVID-19 vaccination did not affect sperm quality and fertilization capacity of men undergoing ART treatments and should be considered safe for men's reproductive health.

Epigenetic Immune Remodeling of Mesothelioma Cells: A New Strategy to Improve the Efficacy of Immunotherapy.

Malignant pleural mesothelioma (MPM) is an aggressive malignancy with a severe prognosis, and with a long-standing need for more effective therapeutic approaches. However, treatment with immune checkpoint inhibitors is becoming an increasingly effective strategy for MPM patients. In this scenario, epigenetic modifications may negatively regulate the interplay between immune and malignant cells within the tumor microenvironment, thus contributing to the highly immunosuppressive contexture of MPM that may limit the efficacy of immunotherapy. Aiming to further improve prospectively the clinical efficacy of immunotherapeutic approaches in MPM, we investigated the immunomodulatory potential of different classes of epigenetic drugs (i.e., DNA hypomethylating agent (DHA) guadecitabine, histone deacetylase inhibitors VPA and SAHA, or EZH2 inhibitors EPZ-6438) in epithelioid, biphasic, and sarcomatoid MPM cell lines, by cytofluorimetric and real-time PCR analyses. We also characterized the effects of the DHA, guadecitabine, on the gene expression profiles (GEP) of the investigated MPM cell lines by the nCounter platform. Among investigated drugs, exposure of MPM cells to guadecitabine, either alone or in combination with VPA, SAHA and EPZ-6438 demonstrated to be the main driver of the induction/upregulation of immune molecules functionally crucial in host-tumor interaction (i.e., HLA class I, ICAM-1 and cancer testis antigens) in all three MPM subtypes investigated. Additionally, GEP demonstrated that treatment with guadecitabine led to the activation of genes involved in several immune-related functional classes mainly in the sarcomatoid subtype. Furthermore, among investigated MPM subtypes, DHA-induced CDH1 expression that contributes to restoring the epithelial phenotype was highest in sarcomatoid cells. Altogether, our results contribute to providing the rationale to develop new epigenetically-based immunotherapeutic approaches for MPM patients, potentially tailored to the specific histologic subtypes.

Immunotherapy of brain metastases: breaking a "dogma".

Until very few years ago, the oncology community dogmatically excluded any clinical potential for immunotherapy in controlling brain metastases. Therefore, despite the significant therapeutic efficacy of monoclonal antibodies to immune check-point(s) across a wide range of tumor types, patients with brain disease were invariably excluded from clinical trials with these agents. Recent insights on the immune landscape of the central nervous system, as well as of the brain tumor microenvironment, are shedding light on the immune-biology of brain metastases.Interestingly, retrospective analyses, case series, and initial prospective clinical trials have recently investigated the role of different immune check-point inhibitors in brain metastases, reporting a significant clinical activity also in this subset of patients. These findings, and their swift translation in the daily practice, are driving fundamental changes in the clinical management of patients with brain metastases, and raise important neuroradiologic challenges. Along this line, neuro-oncology undoubtedly represents an additional area of active investigation and of growing interest to support medical oncologists in the evaluation of clinical responses of brain metastases to ICI treatment, and in the management of neurologic immune-related adverse events.Aim of this review is to summarize the most recent findings on brain metastases immunobiology, on the evolving scenario of clinical efficacy of ICI therapy in patients with brain metastases, as well as on the increasing relevance of neuroradiology in this therapeutic setting.