The role and participation of immune cells in the endometrial tumor microenvironment.

The tumor microenvironment is surrounded by blood vessels and consists of malignant, non-malignant, and immune cells, as well as signalling molecules, which primarily affect the therapeutic response and curative effects of drugs in clinical studies. Tumor-infiltrating immune cells participate in tumor progression, impact anticancer therapy, and eventually lead to the development of immune tolerance. Immunotherapy is evolving as a promising therapeutic intervention to stimulate and activate the immune system to suppress cancer cell growth. Endometrial cancer (EC) is an immunogenic disease, and in recent years, immunotherapy has shown benefit in the treatment of recurrent and advanced EC. This review discusses the key molecular pathways associated with the intra-tumoral immune response and the involvement of circulatory signalling molecules. Specific immunologic signatures in EC which offer targets for immunomodulating agents, are also discussed. We have summarized the available literature in support of using immunotherapy in EC. Lastly, we have also discussed ongoing clinical trials that may offer additional promising immunotherapy options in the future. The manuscript also explored innovative approaches for screening and identifying effective drugs, and to reduce the financial burdens for the development of personalized treatment strategies. Collectively, we aim to provide a comprehensive review of the role of immune cells and the tumor microenvironment in the development, progression, and treatment of EC.

Malaria Biomimetic for Tumor Targeted Drug Delivery.

Malaria infected erythrocytes utilize the parasite protein VAR2CSA to bind to a unique presentation of chondroitin sulfate (CS) for their placenta specific tropism. Interestingly, many cancers express a similar form of CS, thereby termed oncofetal CS (ofCS). The distinctive tropism of malaria infected erythrocytes and the identification of oncofetal CS, therefore, represent potentially potent tools for cancer targeting. Here we describe an intriguing drug delivery platform that effectively mimics infected erythrocytes and their specificity for ofCS. We used a lipid catcher-tag conjugation system for the functionalization of erythrocyte membrane-coated drug carriers with recombinant VAR2CSA (rVAR2). We show that these malaria mimicking erythrocyte nanoparticles (MMENPs) loaded with docetaxel (DTX) specifically target and kill melanoma cells in vitro. We further demonstrate effective targeting and therapeutic efficacy in a xenografted melanoma model. These data thus provide a proof of concept for the use of a malaria biomimetic for tumor targeted drug delivery. Given the broad presentation of ofCS found across various types of malignancies, this biomimetic may therefore show potential as a broadly targeted cancer therapy against multiple tumor indications.

Bispecific T cell-engager targeting oncofetal chondroitin sulfate induces complete tumor regression and protective immune memory in mice.

BACKGROUND: The malaria protein VAR2CSA binds oncofetal chondroitin sulfate (ofCS), a unique chondroitin sulfate, expressed on almost all mammalian cancer cells. Previously, we produced a bispecific construct targeting ofCS and human T cells based on VAR2CSA and anti-CD3 (V-aCD3Hu). V-aCD3Hu showed efficacy against xenografted tumors in immunocompromised mice injected with human immune cells at the tumor site. However, the complex effects potentially exerted by the immune system as a result of the treatment cannot occur in mice without an immune system. Here we investigate the efficacy of V-aCD3Mu as a monotherapy and combined with immune checkpoint inhibitors in mice with a fully functional immune system. METHODS: We produced a bispecific construct consisting of a recombinant version of VAR2CSA coupled to an anti-murine CD3 single-chain variable fragment. Flow cytometry and ELISA were used to check cell binding capabilities and the therapeutic effect was evaluated in vitro in a killing assay. The in vivo efficacy of V-aCD3Mu was then investigated in mice with a functional immune system and established or primary syngeneic tumors in the immunologically "cold" 4T1 mammary carcinoma, B16-F10 malignant melanoma, the pancreatic KPC mouse model, and in the immunologically "hot" CT26 colon carcinoma model. RESULTS: V-aCD3Mu had efficacy as a monotherapy, and the combined treatment of V-aCD3Mu and an immune checkpoint inhibitor showed enhanced effects resulting in the complete elimination of solid tumors in the 4T1, B16-F10, and CT26 models. This anti-tumor effect was abscopal and accompanied by a systemic increase in memory and activated cytotoxic and helper T cells. The combined treatment also led to a higher percentage of memory T cells in the tumor without an increase in regulatory T cells. In addition, we observed partial protection against re-challenge in a melanoma model and full protection in a breast cancer model. CONCLUSIONS: Our findings suggest that V-aCD3Mu combined with an immune checkpoint inhibitor renders immunologically "cold" tumors "hot" and results in tumor elimination. Taken together, these data provide proof of concept for the further clinical development of V-aCD3 as a broad cancer therapy in combination with an immune checkpoint inhibitor.

Cannabis use and its relationship with bipolar disorder: A systematic review and meta-analysis.

Cannabis use has been stated as a causal risk factor for the occurrence of schizophrenia and other psychotic disorders. There is a dearth of literature stating the association of cannabis with bipolar disorder. This review aimed to find the repercussion of cannabis use on the onset of the first episode of bipolar disorder and the worsening of the symptoms in pre-existing illness. A thorough systematic review of the existing literature was carried out using the PRISMA guidelines. PubMed, Medline, EMBASE, SCOPUS, and Google-scholar databases were searched for studies fitting our study's inclusion and exclusion criteria. A total of 25 studies were included in the systematic review and out of these 25 studies, five prospective studies met the inclusion criteria for the primary outcome meta-analysis. A total sample of 13,624 individuals was included in these five studies. A fixed effect model was used in the meta-analysis of these five studies and it revealed an association between cannabis and bipolar disorder with an effect size of 2.63 (95% CI: 1.95-3.53) (heterogeneity: chi² = 3.01, df = 3 (P = 0.39); I² = 0%). Our findings propose that cannabis use may precipitate or worsen bipolar disorder. This highlights the importance of the detrimental effect of cannabis use on bipolar disorder and the need to discourage cannabis use in the youth culture. High-quality prospective studies are required to delineate the effect of cannabis use on bipolar disorder.

Cryo-EM reveals the conformational epitope of human monoclonal antibody PAM1.4 broadly reacting with polymorphic malarial protein VAR2CSA.

Malaria during pregnancy is a major global health problem caused by infection with Plasmodium falciparum parasites. Severe effects arise from the accumulation of infected erythrocytes in the placenta. Here, erythrocytes infected by late blood-stage parasites adhere to placental chondroitin sulphate A (CS) via VAR2CSA-type P. falciparum erythrocyte membrane protein 1 (PfEMP1) adhesion proteins. Immunity to placental malaria is acquired through exposure and mediated through antibodies to VAR2CSA. Through evolution, the VAR2CSA proteins have diversified in sequence to escape immune recognition but retained their overall macromolecular structure to maintain CS binding affinity. This structural conservation may also have allowed development of broadly reactive antibodies to VAR2CSA in immune women. Here we show the negative stain and cryo-EM structure of the only known broadly reactive human monoclonal antibody, PAM1.4, in complex with VAR2CSA. The data shows how PAM1.4's broad VAR2CSA reactivity is achieved through interactions with multiple conserved residues of different sub-domains forming conformational epitope distant from the CS binding site on the VAR2CSA core structure. Thus, while PAM1.4 may represent a class of antibodies mediating placental malaria immunity by inducing phagocytosis or NK cell-mediated cytotoxicity, it is likely that broadly CS binding-inhibitory antibodies target other epitopes at the CS binding site. Insights on both types of broadly reactive monoclonal antibodies may aid the development of a vaccine against placental malaria.

Psychological Impact of the COVID-19 Pandemic and Associated Lockdown on Outpatients With Psychiatric Illness: A Cross-Sectional Study.

Objective: To explore the psychological impact of the coronavirus disease 2019 (COVID-19) pandemic and associated lockdown on patients with psychiatric illness.Methods: An online survey-based cross-sectional study was conducted among patients receiving follow-up treatment at a tertiary care center from January to March 2020. The data were collected using a questionnaire about the possible challenges in 3 broad areas: treatment-related challenges, psychosocial difficulties, and concerns related to COVID-19.Results: The majority of patients (72.6%) reported a positive impact due to the increased availability of family support. Patients with depression and anxiety disorders (39.0%) experienced a more negative impact compared to those with psychotic disorders. Many of the psychiatric patients (22.6%) stopped medications and had difficulties accessing health services. Patients also experienced increased interpersonal conflict, sleep difficulties, and a surge in screen time.Conclusions: The findings highlight the difficulties faced by patients with psychiatric illnesses and emphasize the importance of family cohesion during times of crisis.

Reformation of the chondroitin sulfate glycocalyx enables progression of AR-independent prostate cancer.

Lineage plasticity of prostate cancer is associated with resistance to androgen receptor (AR) pathway inhibition (ARPI) and supported by a reactive tumor microenvironment. Here we show that changes in chondroitin sulfate (CS), a major glycosaminoglycan component of the tumor cell glycocalyx and extracellular matrix, is AR-regulated and promotes the adaptive progression of castration-resistant prostate cancer (CRPC) after ARPI. AR directly represses transcription of the 4-O-sulfotransferase gene CHST11 under basal androgen conditions, maintaining steady-state CS in prostate adenocarcinomas. When AR signaling is inhibited by ARPI or lost during progression to non-AR-driven CRPC as a consequence of lineage plasticity, CHST11 expression is unleashed, leading to elevated 4-O-sulfated chondroitin levels. Inhibition of the tumor cell CS glycocalyx delays CRPC progression, and impairs growth and motility of prostate cancer after ARPI. Thus, a reactive CS glycocalyx supports adaptive survival and treatment resistance after ARPI, representing a therapeutic opportunity in patients with advanced prostate cancer.

Internalization and trafficking of CSPG-bound recombinant VAR2CSA lectins in cancer cells.

Proteoglycans are proteins that are modified with glycosaminoglycan chains. Chondroitin sulfate proteoglycans (CSPGs) are currently being exploited as targets for drug-delivery in various cancer indications, however basic knowledge on how CSPGs are internalized in tumor cells is lacking. In this study we took advantage of a recombinant CSPG-binding lectin VAR2CSA (rVAR2) to track internalization and cell fate of CSPGs in tumor cells. We found that rVAR2 is internalized into cancer cells via multiple internalization mechanisms after initial docking on cell surface CSPGs. Regardless of the internalization pathway used, CSPG-bound rVAR2 was trafficked to the early endosomes in an energy-dependent manner but not further transported to the lysosomal compartment. Instead, internalized CSPG-bound rVAR2 proteins were secreted with exosomes to the extracellular environment in a strictly chondroitin sulfate-dependent manner. In summary, our work describes the cell fate of rVAR2 proteins in tumor cells after initial binding to CSPGs, which can be further used to inform development of rVAR2-drug conjugates and other therapeutics targeting CSPGs.

Oral Health Status in Mentally Disabled Children, Dental Care Knowledge of Parents, and the Impact of Audiovisual Oral Health Education Program.

Aims and objectives: The aims and objectives of this study were to evaluate the oral health status in a group of mentally disabled children, to identify the oral health knowledge of parents and to assess the impact of audiovisual oral health education program. Materials and methods: The study sample comprised of 120 mentally disabled subjects aged 5-16 years and 40 parents attending a vocational institute in Jaipur, India. Caries status and oral hygiene status were assessed by DMFT Index and Simplified Oral Hygiene Index (OHI-S) of Greene and Vermillion, respectively. Twenty pre- and post-structured close-ended questions were designed and a 5-minute video on oral health education was prepared. Data was analyzed by using one way ANOVA test followed by Post hoc Tukeys HSD Test, Wilcoxon Signed Rank Test, and Chi-square test. Effectiveness index was calculated to find the impact of AV Teaching Program. Results: Oral hygiene status deteriorates as the severity of mental retardation increases but it was not found significantly associated with oral hygiene status. An effectiveness index of at least 0.5 indicates that the instructional media was satisfactorily effective. Conclusion: Maximum children with poor oral hygiene were in the category of severe mental retardation and mean OHI-S score decreases from severe-to-mild mental retardation. The newly designed audiovisual aid on complete oral health care was effective at some levels in improving the oral health knowledge of the parents/caregivers of mentally retarded children. How to cite this article: Shah RK, Choudhary S, Tandon S. Oral Health Status in Mentally Disabled Children, Dental Care Knowledge of Parents, and the Impact of Audiovisual Oral Health Education Program. Int J Clin Pediatr Dent 2022;15(2):143-148.

Pathways of care and attitudes toward psychotropics in patients with depressive disorders and psychotic disorders.

Objectives: Delay in institution of appropriate mental healthcare is closely linked with attitudes, stigma, and availability of services, which further directs the help seeking pathways. This study was aimed to assess the pathway of care and attitudes toward psychotropic medications among patients with depressive disorder and psychotic disorders. Materials and Methods: Eighty outpatients with depressive disorder and 40 patients with psychotic disorder were assessed in a tertiary care center for pathways to care and attitude toward psychotropics through Attitude toward Psychotropic Medications Questionnaire and Drug attitude inventory-10. Results: The psychiatrist remained the most common first treatment contact (46% in depression and 62% in psychosis). Greater number of patients in psychotic disorder group had first treatment contact with faith healers or exorcist (17.5%), compared to depressive disorder (6.2%). Patients in depressive group had more favorable attitude toward psychotropics compared to psychosis group. Majority of patients had favorable attitude toward psychotropic medications, but they also had substantial misconceptions about side effects, utility, and need of taking lesser than prescribed doses. Conclusions: Although majority of patients had favorable attitude, they also had substantial misconceptions about medications. These issues need to be addressed for better delivery of comprehensive mental healthcare.

Detection of VAR2CSA-Captured Colorectal Cancer Cells from Blood Samples by Real-Time Reverse Transcription PCR.

Analysis of circulating tumor cells (CTCs) from blood samples provides a non-invasive approach for early cancer detection. However, the rarity of CTCs makes it challenging to establish assays with the required sensitivity and specificity. We combine a highly sensitive CTC capture assay exploiting the cancer cell binding recombinant malaria VAR2CSA protein (rVAR2) with the detection of colon-related mRNA transcripts (USH1C and CKMT1A). Cancer cell transcripts are detected by RT-qPCR using proprietary Target Enrichment Long-probe Quantitative Amplified Signal (TELQAS) technology. We validate each step of the workflow using colorectal cancer (CRC) cell lines spiked into blood and compare this with antibody-based cell detection. USH1C and CKMT1A are expressed in healthy colon tissue and CRC cell lines, while only low-level expression can be detected in healthy white blood cells (WBCs). The qPCR reaction shows a near-perfect amplification efficiency for all primer targets with minimal interference of WBC cDNA. Spike-in of 10 cancer cells in 3 mL blood can be detected and statistically separated from control blood using the RT-qPCR assay after rVAR2 capture (p < 0.01 for both primer targets, Mann-Whitney test). Our results provide a validated workflow for highly sensitive detection of magnetically enriched cancer cells.

The specificity of the malarial VAR2CSA protein for chondroitin sulfate depends on 4-O-sulfation and ligand accessibility.

Placental malaria infection is mediated by the binding of the malarial VAR2CSA protein to the placental glycosaminoglycan, chondroitin sulfate. Recombinant subfragments of VAR2CSA (rVAR2) have also been shown to bind specifically and with high affinity to cancer cells and tissues, suggesting the presence of a shared type of oncofetal chondroitin sulfate (ofCS) in the placenta and in tumors. However, the exact structure of ofCS and what determines the selective tropism of VAR2CSA remains poorly understood. In this study, ofCS was purified by affinity chromatography using rVAR2 and subjected to detailed structural analysis. We found high levels of N-acetylgalactosamine 4-O-sulfation (∼80-85%) in placenta- and tumor-derived ofCS. This level of 4-O-sulfation was also found in other tissues that do not support parasite sequestration, suggesting that VAR2CSA tropism is not exclusively determined by placenta- and tumor-specific sulfation. Here, we show that both placenta and tumors contain significantly more chondroitin sulfate moieties of higher molecular weight than other tissues. In line with this, CHPF and CHPF2, which encode proteins required for chondroitin polymerization, are significantly upregulated in most cancer types. CRISPR/Cas9 targeting of CHPF and CHPF2 in tumor cells reduced the average molecular weight of cell-surface chondroitin sulfate and resulted in a marked reduction of rVAR2 binding. Finally, utilizing a cell-based glycocalyx model, we showed that rVAR2 binding correlates with the length of the chondroitin sulfate chains in the cellular glycocalyx. These data demonstrate that the total amount and cellular accessibility of chondroitin sulfate chains impact rVAR2 binding and thus malaria infection.

Social cognition in siblings of patients with bipolar disorders.

BACKGROUND: Identifying people at risk of developing bipolar disorder (BD) using endophenotypes is of recent interest. Few studies on social cognition in first-degree relatives of patients with BD have shown inconsistent findings. This study aimed to evaluate the social cognitive deficits (SCD) and its correlates among siblings of patients with BD. METHODOLOGY: In this cross-sectional study, patients of BD (n = 32), their siblings (n = 32), and healthy control (HC) subjects (n = 38) matched for age, gender, and education were evaluated on social cognition rating tools in Indian setting and neurocognitive tests (color trail test, Wisconsin card sorting test [WCST], and Hopkin's verbal learning test [HVLT]). RESULTS: When the siblings of patients with BD were compared with patients with BD and the HCs, siblings (mean 0.6 [standard deviation [SD]: 0.2]) performed worse than the HCs (mean 0.9 [SD: 0.1]) (P ≤ 0.001) on the Faux pas composite index. Compared to HCs, siblings performed worse on all the subtests of HVLT (Trial delayed) (P < 0.001) and WCST (total correct, total errors, and conceptual responses) (P < 0.001). CONCLUSION: Presence of elevated level of SCD among siblings, especially the Faux pas composite index in comparison to HCs, imply that these are stable traits, which are more often present in the patients and at risk individuals. This implies that SCD can be considered as another important endophenotype for BD.

Oncofetal Chondroitin Sulfate Is a Highly Expressed Therapeutic Target in Non-Small Cell Lung Cancer.

Broad-spectrum therapeutics in non-small cell lung cancer (NSCLC) are in demand. Most human solid tumors express proteoglycans modified with distinct oncofetal chondroitin sulfate (CS) chains that can be detected and targeted with recombinant VAR2CSA (rVAR2) proteins and rVAR2-derived therapeutics. Here, we investigated expression and targetability of oncofetal CS expression in human NSCLC. High oncofetal CS expression is associated with shorter disease-free survival and poor overall survival of clinically annotated stage I and II NSCLC patients (n = 493). Oncofetal CS qualifies as an independent prognosticator of NSCLC in males and smokers, and high oncofetal CS levels are more prevalent in EGFR/KRAS wild-type cases, as compared to mutation cases. NSCLC cell lines express oncofetal CS-modified proteoglycans that can be specifically detected and targeted by rVAR2 proteins in a CSA-dependent manner. Importantly, a novel VAR2-drug conjugate (VDC-MMAE) efficiently eliminates NSCLC cells in vitro and in vivo. In summary, oncofetal CS is a prognostic biomarker and an actionable glycosaminoglycan target in NSCLC.

Cryo-EM reveals the architecture of placental malaria VAR2CSA and provides molecular insight into chondroitin sulfate binding.

Placental malaria can have severe consequences for both mother and child and effective vaccines are lacking. Parasite-infected red blood cells sequester in the placenta through interaction between parasite-expressed protein VAR2CSA and the glycosaminoglycan chondroitin sulfate A (CS) abundantly present in the intervillous space. Here, we report cryo-EM structures of the VAR2CSA ectodomain at up to 3.1 Å resolution revealing an overall V-shaped architecture and a complex domain organization. Notably, the surface displays a single significantly electropositive patch, compatible with binding of negatively charged CS. Using molecular docking and molecular dynamics simulations as well as comparative hydroxyl radical protein foot-printing of VAR2CSA in complex with placental CS, we identify the CS-binding groove, intersecting with the positively charged patch of the central VAR2CSA structure. We identify distinctive conserved structural features upholding the macro-molecular domain complex and CS binding capacity of VAR2CSA as well as divergent elements possibly allowing immune escape at or near the CS binding site. These observations will support rational design of second-generation placental malaria vaccines.

Development of a bispecific immune engager using a recombinant malaria protein.

As an immune evasion and survival strategy, the Plasmodium falciparum malaria parasite has evolved a protein named VAR2CSA. This protein mediates sequestration of infected red blood cells in the placenta through the interaction with a unique carbohydrate abundantly and exclusively present in the placenta. Cancer cells were found to share the same expression of this distinct carbohydrate, termed oncofetal chondroitin sulfate on their surface. In this study we have used a protein conjugation system to produce a bispecific immune engager, V-aCD3, based on recombinant VAR2CSA as the cancer targeting moiety and an anti-CD3 single-chain variable fragment linked to a single-chain Fc as the immune engager. Conjugation of these two proteins resulted in a single functional moiety that induced immune mediated killing of a broad range of cancer cells in vitro and facilitated tumor arrest in an orthotopic bladder cancer xenograft model.

Capsid-like particles decorated with the SARS-CoV-2 receptor-binding domain elicit strong virus neutralization activity.

The rapid development of a SARS-CoV-2 vaccine is a global priority. Here, we develop two capsid-like particle (CLP)-based vaccines displaying the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. RBD antigens are displayed on AP205 CLPs through a split-protein Tag/Catcher, ensuring unidirectional and high-density display of RBD. Both soluble recombinant RBD and RBD displayed on CLPs bind the ACE2 receptor with nanomolar affinity. Mice are vaccinated with soluble RBD or CLP-displayed RBD, formulated in Squalene-Water-Emulsion. The RBD-CLP vaccines induce higher levels of serum anti-spike antibodies than the soluble RBD vaccines. Remarkably, one injection with our lead RBD-CLP vaccine in mice elicits virus neutralization antibody titers comparable to those found in patients that had recovered from COVID-19. Following booster vaccinations, the virus neutralization titers exceed those measured after natural infection, at serum dilutions above 1:10,000. Thus, the RBD-CLP vaccine is a highly promising candidate for preventing COVID-19.

Relapse in bipolar disorder in hospitalized patients with COVID-19: A case series and key recommendations.

COVID-19 has adversely affected the lives of people with bipolar disorder (BD). We have reported three patients with BD, who suffered from relapse after being diagnosed with COVID-19. Patient X is a 54-year-old man who developed manic episode, while Y is a 63-year-old man and Z is a 52-year-old woman who developed severe depression after being hospitalized for COVID-19. In this case series, three patients with BD who developed manic or depressive episodes during COVID-19 infection have been described. Adequate liaison between consultation-liaison psychiatrist and primary medical team was done. Rational pharmacotherapy along with psychosocial interventions led to favorable outcomes in all three cases.