UMG1/CD3ε-bispecific T-cell engager redirects T-cell cytotoxicity against diffuse large B-cell lymphoma.

UMG1 is a unique epitope of CD43, not expressed by normal cells and tissues of haematopoietic and non-haematopoietic origin, except thymocytes and a minority (<5%) of peripheral blood T lymphocytes. By immunohistochemistry analysis of tissue microarray and pathology slides, we found high UMG1 expression in 20%-24% of diffuse large B-cell lymphomas (DLBCLs), including highly aggressive BCL2high and CD20low cases. UMG1 membrane expression was also found in DLBCL bone marrow-infiltrating cells and established cell lines. Targeting UMG1 with a novel asymmetric UMG1/CD3ε-bispecific T-cell engager (BTCE) induced redirected cytotoxicity against DLBCL cells and was synergistic with lenalidomide. We conclude that UMG1/CD3ε-BTCE is a promising therapeutic for DLBCLs.

Victim Blaming: Being a Victim Twice. Comparison of Emotional and Socio-Cultural Aspects.

World Health Organization Director-General Margaret Chan said that violence against women is a global structural issue. It is a health problem of epidemic dimensions, the analysis of 141 researches carried out in 81 countries shows that 35% of women suffer some form of violence during their lifetime. These data were presented in the largest study ever done on the physical and sexual abuse suffered by women in all regions of the planet. When we talk about violence against women we refer to the definition provided by the United Nations and the World Health Organization. As regards the definition provided by the UN, the expression covers "any act of gender-based violence that results in, or is likely to result in, physical, sexual or psychological harm or suffering, including threats of such acts, coercion and arbitrary deprivation of liberty, both in public and private life". The WHO definition outlines violence as "the intentional use of physical force or power, threatened or actual, against oneself. But the definitions don't end there. The Convention of the Council of Europe, the first regulatory element on the matter of preventing and combating violence against women, states that the term "domestic violence" refers to all acts of violence, whether physical, sexual, psychological or economic, occurring within the family or household or between current or former spouses or partners, whether or not the perpetrator shares or has shared the same residence with the victim. The document, which starts from the same cultural reading of the roots of violence against women, commits the signatory states to protect women from all forms of violence and to prevent, prosecute and eliminate violence against women and domestic violence. The Convention requires states parties to organize "specialised immediate, short- and long-term support services for each victim of any act of violence falling within the scope" of the Convention. Unfortunately violence is a dynamic event, in fact it is constantly changing, so we are witnessing a continuous transformation of the forms of violence. In particular, in this study we will address the issue of secondary victimization, which unfortunately represents an increasingly present form of violence. citation. This form of violence was defined by the American psychologist, William J. Ryan, Jr, who in 1971 defined it as a phenomenon of secondary victimization.

Gender Dysphoria and DNA.

Gender dysphoria (GD) describes individuals for whom the native sex and expressed gender are not coincident and most of them self-identify as transgender women or men. It has been shown that genetic factors play an important role in GD and the presence of specific genetic variants in candidate genes could be correlated. On the other hand, twins studies have estimated its heritability. In this review, we collect and report the available data obtained by different molecular genetic studies.

Eating Disorders and Psychiatric Comorbidity.

Psychiatric comorbidity is present in more than 70% of people with an Eating Disorders (ED), before or during the acute state of illness or in the long-term course. These comorbidities include personality disorders (>53%), anxiety disorders (>50%), mood disorders (>40%) and substance abuse (>10%). This work aims to analyse the different treatments available for patients affected by eating disorders and other psychiatric comorbidity.

Adolescents and the Consequences of COVID-19.

In the pandemic period due to the strong restrictions made necessary to deal with the spread of the Sars COVID-19 Virus, adolescents were the subjects who, most of all, suffered from isolation and lack of sociality, due to social distancing. It was necessary to change their habits, their lifestyles, the way they live social relationships and relate to others. Furthermore, those of them who were facing health difficulties at the time were faced with the impossibility of the services to guarantee safety conditions, so that in most cases many medical, psychiatric and psychological activities were suspended, with hospitalizations postponed to a date to be defined, appointments canceled with a high risk of losing contact with the patient. Many authors agreed, right from the start, that the pandemic consequences would have focused above all on psychological problems. The scientific literature on past pandemics had shown the role of fear, anxiety, stress and depression and its negative psychosocial consequences on the quality of life of the population. Samantha Brooks and colleagues from King's College London reviewed 24 studies on the psychological impact of quarantine and highlighted negative psychological effects including symptoms of post-traumatic stress, confusion and anger. Today there are many contributions in the literature which unfortunately confirm what has been hypothesized.

Hit identification of novel small molecules interfering with MALAT1 triplex by a structure-based virtual screening.

Nowadays, RNA is an attractive target for the design of new small molecules with different pharmacological activities. Among several RNA molecules, long noncoding RNAs (lncRNAs) are extensively reported to be involved in cancer pathogenesis. In particular, the overexpression of lncRNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) plays an important role in the development of multiple myeloma (MM). Starting from the crystallographic structure of the triple-helical stability element at the 3'-end of MALAT1, we performed a structure-based virtual screening of a large commercial database, previously filtered according to the drug-like properties. After a thermodynamic analysis, we selected five compounds for the in vitro assays. Compound M5, characterized by a diazaindene scaffold, emerged as the most promising molecule enabling the destabilization of the MALAT1 triplex structure and antiproliferative activity on in vitro models of MM. M5 is proposed as a lead compound to be further optimized for improving its affinity toward MALAT1.

A Pronectin™ AXL-targeted first-in-class bispecific T cell engager (pAXLxCD3ε) for ovarian cancer.

BACKGROUND: Pronectins™ are a new class of fibronectin-3-domain 14th-derived (14Fn3) antibody mimics that can be engineered as bispecific T cell engager (BTCE) to redirect immune effector cells against cancer. We describe here the in vitro and in vivo activity of a Pronectin™ AXL-targeted first-in-class bispecific T cell engager (pAXLxCD3ε) against Epithelial Ovarian Cancer (EOC). METHODS: pAXLxCD3ε T-cell mediated cytotoxicity was evaluated by flow cytometry and bioluminescence. pAXLxCD3ε mediated T-cell infiltration, activation and proliferation were assessed by immunofluorescence microscopy and by flow cytometry. Activity of pAXLxCD3ε was also investigated in combination with poly-ADP ribose polymerase inhibitors (PARPi). In vivo antitumor activity of pAXLxCD3ε was evaluated in immunocompromised (NSG) mice bearing intraperitoneal or subcutaneous EOC xenografts and immunologically reconstituted with human peripheral blood mononuclear cells (PBMC). RESULTS: pAXLxCD3ε induced dose-dependent cytotoxicity by activation of T lymphocytes against EOC cells, regardless of their histologic origin. The addition of PARPi to cell cultures enhanced pAXLxCD3ε cytotoxicity. Importantly, in vivo, pAXLxCD3ε was highly effective against EOC xenografts in two different NSG mouse models, by inhibiting the growth of tumor cells in ascites and subcutaneous xenografts. This effect translated into a significantly prolonged survival of treated animals. CONCLUSION: pAXLxCD3ε is an active therapeutics against EOC cells providing a rational for its development as a novel agent in this still incurable disease. The preclinical validation of a first-in-class agent opens the way to the development of a new 14Fn3-based scaffold platform for the generation of innovative immune therapeutics against cancer.

TERRA G-quadruplex stabilization as a new therapeutic strategy for multiple myeloma.

BACKGROUND: Multiple myeloma (MM) is a hematologic malignancy characterized by high genomic instability, and telomere dysfunction is an important cause of acquired genomic alterations. Telomeric repeat-containing RNA (TERRA) transcripts are long non-coding RNAs involved in telomere stability through the interaction with shelterin complex. Dysregulation of TERRAs has been reported across several cancer types. We recently identified a small molecule, hit 17, which stabilizes the secondary structure of TERRA. In this study, we investigated in vitro and in vivo anti-MM activities of hit 17. METHODS: Anti-proliferative activity of hit 17 was evaluated in different MM cell lines by cell proliferation assay, and the apoptotic process was analyzed by flow cytometry. Gene and protein expressions were detected by RT-qPCR and western blotting, respectively. Microarray analysis was used to analyze the transcriptome profile. The effect of hit 17 on telomeric structure was evaluated by chromatin immunoprecipitation. Further evaluation in vivo was proceeded upon NCI-H929 and AMO-1 xenograft models. RESULTS: TERRA G4 stabilization induced in vitro dissociation of telomeric repeat-binding factor 2 (TRF2) from telomeres leading to the activation of ATM-dependent DNA damage response, cell cycle arrest, proliferation block, and apoptotic death in MM cell lines. In addition, up-regulation of TERRA transcription was observed upon DNA damage and TRF2 loss. Transcriptome analysis followed by gene set enrichment analysis (GSEA) confirmed the involvement of the above-mentioned processes and other pathways such as E2F, MYC, oxidative phosphorylation, and DNA repair genes as early events following hit 17-induced TERRA stabilization. Moreover, hit 17 exerted anti-tumor activity against MM xenograft models. CONCLUSION: Our findings provide evidence that targeting TERRA by hit 17 could represent a promising strategy for a novel therapeutic approach to MM.

The First-In-Class Anti-AXL×CD3ε Pronectin™-Based Bispecific T-Cell Engager Is Active in Preclinical Models of Human Soft Tissue and Bone Sarcomas.

Sarcomas are heterogeneous malignancies with limited therapeutic options and a poor prognosis. We developed an innovative immunotherapeutic agent, a first-in-class Pronectin™-based Bispecific T-Cell Engager (pAXL×CD3ε), for the targeting of AXL, a TAM family tyrosine kinase receptor highly expressed in sarcomas. AXL expression was first analyzed by flow cytometry, qRT-PCR, and Western blot on a panel of sarcoma cell lines. The T-cell-mediated pAXL×CD3ε cytotoxicity against sarcoma cells was investigated by flow cytometry, luminescence assay, and fluorescent microscopy imaging. The activation and degranulation of T cells induced by pAXL×CD3ε were evaluated by flow cytometry. The antitumor activity induced by pAXL×CD3ε in combination with trabectedin was also investigated. In vivo activity studies of pAXL×CD3ε were performed in immunocompromised mice (NSG), engrafted with human sarcoma cells and reconstituted with human peripheral blood mononuclear cells from healthy donors. Most sarcoma cells showed high expression of AXL. pAXL×CD3ε triggered T-lymphocyte activation and induced dose-dependent T-cell-mediated cytotoxicity. The combination of pAXL×CD3ε with trabectedin increased cytotoxicity. pAXL×CD3ε inhibited the in vivo growth of human sarcoma xenografts, increasing the survival of treated mice. Our data demonstrate the antitumor efficacy of pAXL×CD3ε against sarcoma cells, providing a translational framework for the clinical development of pAXL×CD3ε in the treatment of human sarcomas, aggressive and still-incurable malignancies.

Exploiting DNA Ligase III addiction of multiple myeloma by flavonoid Rhamnetin.

BACKGROUND: DNA ligases are crucial for DNA repair and cell replication since they catalyze the final steps in which DNA breaks are joined. DNA Ligase III (LIG3) exerts a pivotal role in Alternative-Non-Homologous End Joining Repair (Alt-NHEJ), an error-prone DNA repair pathway often up-regulated in genomically unstable cancer, such as Multiple Myeloma (MM). Based on the three-dimensional (3D) LIG3 structure, we performed a computational screening to identify LIG3-targeting natural compounds as potential candidates to counteract Alt-NHEJ activity in MM. METHODS: Virtual screening was conducted by interrogating the Phenol Explorer database. Validation of binding to LIG3 recombinant protein was performed by Saturation Transfer Difference (STD)-nuclear magnetic resonance (NMR) experiments. Cell viability was analyzed by Cell Titer-Glo assay; apoptosis was evaluated by flow cytometric analysis following Annexin V-7AAD staining. Alt-NHEJ repair modulation was evaluated using plasmid re-joining assay and Cytoscan HD. DNA Damage Response protein levels were analyzed by Western blot of whole and fractionated protein extracts and immunofluorescence analysis. The mitochondrial DNA (mtDNA) copy number was determined by qPCR. In vivo activity was evaluated in NOD-SCID mice subcutaneously engrafted with MM cells. RESULTS: Here, we provide evidence that a natural flavonoid Rhamnetin (RHM), selected by a computational approach, counteracts LIG3 activity and killed Alt-NHEJ-dependent MM cells. Indeed, Nuclear Magnetic Resonance (NMR) showed binding of RHM to LIG3 protein and functional experiments revealed that RHM interferes with LIG3-driven nuclear and mitochondrial DNA repair, leading to significant anti-MM activity in vitro and in vivo. CONCLUSION: Taken together, our findings provide proof of concept that RHM targets LIG3 addiction in MM and may represent therefore a novel promising anti-tumor natural agent to be investigated in an early clinical setting.

Therapeutic activation of G protein-coupled estrogen receptor 1 in Waldenström Macroglobulinemia.

Activating G protein-coupled estrogen receptor 1 (GPER1) is an attractive therapeutic strategy for treating a variety of human diseases including cancer. Here, we show that GPER1 is significantly upregulated in tumor cells from different cohorts of Waldenström Macroglobulinemia (WM) patients compared to normal B cells. Using the clinically applicable GPER1-selective small-molecule agonist G-1 (also named Tespria), we found that pharmacological activation of GPER1 leads to G2/M cell cycle arrest and apoptosis both in vitro and in vivo in animal models, even in the context of the protective bone marrow milieu. Activation of GPER1 triggered the TP53 pathway, which remains actionable during WM progression. Thus, this study identifies a novel therapeutic target in WM and paves the way for the clinical development of the GPER1 agonist G-1.

Eating Disorders: The Role of the Family in Development and Maintenance of Children's Problems in the Pandemic Period.

INTRODUCTION: According to data released by the Ministry of Health in 2021 in Italy about three million young people suffer from eating disorders with onset before the age of 13 and the number tends to be increasing. This work aims to understand if and to what extent the areas of family functioning are related to the way of eating of adolescents in the period of restriction due to COVID-19. In particular, which dimensions of family functioning can be correlated with dysfunctional eating habits. METHODOLOGY: The group that took part in the study was composed of 154 non clinical subjects, of which 124 females, 27 males and 3 non-binary gender subjects. The tests used were the McMaster Family Assessment Device and the Binge Eating Scale, in addition a personal data sheet was used containing the details of the subjects who participated anonymously, recruited at the university of Italy. The data have some limitations, first of all the low number of the sample and the online modality in compiling the tests. RESULTS: In general, we can say that the Fad and the Bes correlate positively, as the subscales of the Fad increase, the disorganized eating behavior increases and therefore the score of the Bes test. The results of the study indicate that some family dynamics are related to disorganized eating behaviors; it is not possible to establish the specific gravity of the pandemic. It seems likely that compromised family relationships may play a role in promoting the onset of diseases. CONCLUSION: It is appropriate to think of a psychoeducational intervention aimed at families in order to improve immediate family functioning and guarantee young people effective preventive action in subjects at risk in adolescence.

Violence Against Women: An Ever-Increasing Emergency. Path of Escape from Violence.

Violence against women is a complex phenomenon that poses many difficulties. The greatest obstacles, however, are those that women themselves encounter, often with children, who choose to get out of the situation or relationship of violence they experience. Material, economic, housing, psychological difficulties; in this difficult path to escape from violence, women need support and support, needs to which I respond to the Anti-violence Centers (CAV) that have been operating in the various territories for years. In Italy, in 2021 ISTAT published a survey on women who have found listening, services, accompaniment and protection at the CAVs and shelters operating on the Italian territory: they are 54,609 (those who have contacted the CAVs at least once), 3,964 more than in 2019; 30,359, on the other hand, have started a path to escape from violence with the CAVs that adhere to the Intesa Stato Regioni, of which 20,223 (66.1% of the total number of women taken into care) have started the process in 2020 (69, 1% the previous year). On 13 July 2022 it was published by D.i.Re. (Women on the Net against violence) a specific and detailed report on the collection of data on women received by the CAVs of the same D.i.Re network in which: 20,711 women, 3.5% more contacts than in 2020, 8 8% more women who had never turned to CAVs. The data takes into account what we have experienced following the restrictions dictated by the Covid -19 pandemic, which has seen the growth of requests for help from women and the difficulty of activating the path of escape from violence with particular important repercussions on the victims who could not even go to hospital emergency rooms clogged with coronavirus patients.

Genetics As a Key to Understand Mood Disorders.

Mood disorders are mental health class that health professionals use to broadly describe all types of depression and bipolar disorders. Heritability of both bipolar and depressive disorder is in the range of 50%, which means that genes alone are not sufficient to explain all the cases of mood disorders, but they confer a substantial risk which is combined with environmental stressors to determine the final illness. In the recent years, a number of studies considered the idea to develop a strategic plan to employ the tools of genetics to advance the understanding, treatment, and outcomes for mood disorders.

Eating and Nutrition Disorders: Intra and Post-Lockdown Pandemic Emergencies.

Eating Disorders (ED), currently specified as Eating and Nutrition Disorders (DAN), have impacted their morbidity since the last years of the last century, afflicting large segments of the population, predominantly youth, in the westernized "hemisphere." In addition, the COVID-19 pandemic has impacted and negatively influenced eating behaviors in both the general population and DAN sufferers. In agreement with many authors, distancing and social isolation have produced eating disconducts or aggravated symptoms in patients undergoing treatment. All this has led in many cases, to the demand for urgent and-or emergency care. This paper aims to review recent literature and expose data on such treatment regimens.

The New Microtubule-Targeting Agent SIX2G Induces Immunogenic Cell Death in Multiple Myeloma.

Microtubule-targeting agents (MTAs) are effective drugs for cancer treatment. A novel diaryl [1,2]oxazole class of compounds binding the colchicine site was synthesized as cis-restricted-combretastatin-A-4-analogue and then chemically modified to have improved solubility and a wider therapeutic index as compared to vinca alkaloids and taxanes. On these bases, a new class of tricyclic compounds, containing the [1,2]oxazole ring and an isoindole moiety, has been synthetized, among which SIX2G emerged as improved MTA. Several findings highlighted the ability of some chemotherapeutics to induce immunogenic cell death (ICD), which is defined by the cell surface translocation of Calreticulin (CALR) via dissociation of the PP1/GADD34 complex. In this regard, we computationally predicted the ability of SIX2G to induce CALR exposure by interacting with the PP1 RVxF domain. We then assessed both the potential cytotoxic and immunogenic activity of SIX2G on in vitro models of multiple myeloma (MM), which is an incurable hematological malignancy characterized by an immunosuppressive milieu. We found that the treatment with SIX2G inhibited cell viability by inducing G2/M phase cell cycle arrest and apoptosis. Moreover, we observed the increase of hallmarks of ICD such as CALR exposure, ATP release and phospho-eIF2α protein level. Through co-culture experiments with immune cells, we demonstrated the increase of (i) CD86 maturation marker on dendritic cells, (ii) CD69 activation marker on cytotoxic T cells, and (iii) phagocytosis of tumor cells following treatment with SIX2G, confirming the onset of an immunogenic cascade. In conclusion, our findings provide a framework for further development of SIX2G as a new potential anti-MM agent.

A Novel Bispecific T-Cell Engager (CD1a x CD3ε) BTCE Is Effective against Cortical-Derived T Cell Acute Lymphoblastic Leukemia (T-ALL) Cells.

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy burdened by poor prognosis. While huge progress of immunotherapy has recently improved the outcome of B-cell malignancies, the lack of tumor-restricted T-cell antigens still hampers its progress in T-ALL. Therefore, innovative immunotherapeutic agents are eagerly awaited. To this end, we generated a novel asymmetric (2 + 1) bispecific T-cell engager (BTCE) targeting CD1a and CD3ε (CD1a x CD3ε) starting from the development of a novel mAb named UMG2. UMG2 mAb reacts against CD1a, a glycoprotein highly expressed by cortical T-ALL cells. Importantly, no UMG2 binding was found on normal T-cells. CD1a x CD3ε induced high T-cell mediated cytotoxicity against CD1a+ T-ALL cells in vitro, as demonstrated by the concentration-dependent increase of T-cell proliferation, degranulation, induction of cell surface activation markers, and secretion of pro-inflammatory cytokines. Most importantly, in a PBMC-reconstituted NGS mouse model bearing human T-ALL, CD1a x CD3ε significantly inhibited the growth of human T-ALL xenografts, translating into a significant survival advantage of treated animals. In conclusion, CD1a x CD3ε is a novel BTCE highly active against CD1a-expressing cortical-derived T-ALL cells suitable for clinical development as an effective therapeutic option for this rare and aggressive disease.

Chromene Derivatives as Selective TERRA G-Quadruplex RNA Binders with Antiproliferative Properties.

In mammalian cells, telomerase transcribes telomeres in large G-rich non-coding RNA, known as telomeric repeat-containing RNA (TERRA), which folds into noncanonical nucleic acid secondary structures called G-quadruplexes (G4s). Since TERRA G4 has been shown to be involved in telomere length and translation regulation, it could provide valuable insight into fundamental biological processes, such as cancer growth, and TERRA G4 binders could represent an innovative strategy for cancer treatment. In this work, the three best candidates identified in our previous virtual screening campaign on bimolecular DNA/RNA G4s were investigated on the monomolecular Tel DNA and TERRA G4s by means of molecular modelling simulations and in vitro and in cell analysis. The results obtained in this work highlighted the stabilizing power of all the three candidates on TERRA G4. In particular, the two compounds characterized by a chromene scaffold were selective TERRA G4 binders, while the compound with a naphthyridine core acted as a dual Tel/TERRA G4-binder. A biophysical investigation by circular dichroism confirmed the relative stabilization efficiency of the compounds towards TERRA and Tel G4s. The TERRA G4 stabilizing hits showed good antiproliferative activity against colorectal and lung adenocarcinoma cell lines. Lead optimization to increase TERRA G4 stabilization may provide new powerful tools against cancer.

Oleil Hydroxytyrosol (HTOL) Exerts Anti-Myeloma Activity by Antagonizing Key Survival Pathways in Malignant Plasma Cells.

Polyphenols from olive oil are endowed with several biological activities. Chemical modifications have been recently applied to these compounds to improve their therapeutic activity in different pathological settings, including cancer. Herein, we describe the in vitro effects on multiple myeloma (MM) cells of oleil hydroxytyrosol (HTOL), a synthetic fatty ester of natural hydroxytyrosol with oleic acid. HTOL reduced the viability of various human MM cell lines (HMCLs), even when co-cultured with bone marrow stromal cells, triggering ER stress, UPR and apoptosis, while it was not cytotoxic against healthy peripheral blood mononuclear cells or B lymphocytes. Whole-transcriptome profiling of HTOL-treated MM cells, coupled with protein expression analyses, indicate that HTOL antagonizes key survival pathways for malignant plasma cells, including the undruggable IRF4-c-MYC oncogenic axis. Accordingly, c-MYC gain- and loss-of-function strategies demonstrate that HTOL anti-tumor activity was, at least in part, due to c-MYC targeting. Taken together, these findings underscore the anti-MM potential of HTOL, providing the molecular framework for further investigation of HTOL-based treatments as novel anti-cancer agents.

Endophenotype and Psychiatry: an Interesting Combination.

The endophenotype is a measurable component which is characterized as an intermediate part of the path existing between the genotype and phenotype of a disease. In the context of psychiatric pathologies, an endophenotype is such if it shares inheritable variations with it, if it is evident both during the active and inactive phases of the pathology, if it is co-transmitted in the family, and if it is evident in both affected family members from pathology, and in healthy people. This review reports the available literature data of the interesting combination between endophenotype and psychiatry.