Academic literature on the topic 'Salute transgender'

Il concetto di identità di genere è stato motivo di interesse e di studio negli anni fino a giungere alla sua concezione transgender, caratterizzata da un lungo percorso di depatologizzazione che parte dai primi studi di differenziazione tra sesso e genere fino ad arrivare alle classificazioni che i vari manuali diagnostici oggi ci presentano. La concezione transgender dei generi supera la concezione binaria, proponendo una visione fluida che legittima l'esistenza di una varietà di identità di genere in cui potersi riconoscere. I bisogni relativi all'invecchiamento e alla salute degli anziani LGBT sono scarsamente affrontati nei servizi, nelle politiche o nella ricerca. La popolazione transgender over 50, la quale ha vissuto in un contesto so-ciale e storico caratterizzato da forti discriminazioni, si trova ad affrontare una fase di transizione della vita in cui è importante considerare le caratteristiche peculiari di tale vissuto come transgender e gli aspetti relativi alla salute fisica e al supporto sociale. Affacciarci alla visione di invecchiamento transgender vuol dire adottare una prospettiva che si distanzia dagli script eteronormativi prevalenti nella società, e che si avvale di concezioni tipiche di tale vissuto, come quella di queer time e queer space. L'obiettivo di tale contributo è quello di evidenziare le problematiche e le esigenze della popolazione considerata per poter promuovere una figura di clinico attento e consapevole delle peculiarità dell'utenza considerata e un intervento clinico psicosessuologico ad hoc che adotti un approccio biopsicosociale.

Le ricerche in ambito cardiovascolare evidenziano che la popolazione LGBT+ vive una disparità di salute rispetto alla popolazione coetanea eterosessuale e cisgender. Le minoranze sessuali sono infatti sottoposte a stressors specifici che intervengono a più livelli nella vita della persona e che, in assenza di peculiari fattori protettivi di resilienza, hanno un impatto negativo sulla salute cardiovascolare. Restano inoltre da raccogliere le evidenze degli effetti delle terapie ormonali di affermazione di genere e dei bloccanti ipotalamici sulla salute cardiovascolare, per consentire ai medici e alle persone transgender e gender-diverse di conoscere il rischio a cui si sottopongono e di gestirlo in maniera adeguata. Nonostante le raccomandazioni di più istituti scientifici, attualmente si rileva una condizione di impreparazione e di incertezza del personale sanitario su come approcciare l’utenza LGBT+ dal punto di vista sia clinico sia relazionale. Per ridurre le disparità in ambito di salute cardiovascolare che riguardano gli individui LGBT+, si rivela necessario approfondire le conoscenze sui fattori biologici e psico-sociali che potenziano i fattori di rischio cardiovascolare di questa popolazione, sviluppare linee guida evidence-based per prendersi cura dei bisogni specifici di salute delle minoranze sessuali, e avviare programmi di formazione al personale sanitario volti a ridurre gli atteggiamenti discriminatori e condividere buone pratiche a garanzia di una medicina dell’uguaglianza.

Abstract Retroviral vectors are currently the most suitable vehicles for therapeutic gene transfer in hematopoietic stem cells. However, these vectors are known to integrate rather randomly throughout the genome, suffering the so called chromosomal position effects (PE). Such a critical occurrence most probably depends upon the ability of heterochromatin to spread in the inserted vector sequences. Moreover, the use of transgenes imply genotoxicity effects, since the cis-regulatory sequences harbored by the vector can disturb the proper transcription of the resident genes neighboring the integration site, potentially leading to malignant transformation. Due to their enhancer blocker activity, the incorporation of chromatin insulators in flanking position to the transferred unit can reduce the mentioned dangerous effects. Moreover, by acting as barriers to the spread of heterochromatin, chromatin insulators can also mitigate vector silencing. We have previously shown that the sea urchin sns5 chromatin insulator activity is conserved in mouse and human erythroid milieu: it blocks the βglobin-LCR-HS2 enhancer/globin promoter interaction when placed between them. In addition, when placed in flanking location of a γ-retrovirus vector, sns5 impedes PE variegation and improves vector-specific expression following integration in the erythroid genome. Importantly, by binding both erythroid-specific and ubiquitous factors, sns5 favors the accumulation inside the provirus locus of epigenetic marks commonly associated to an euchromatic state (Acuto S. et al., BCMD 2005; D'Apolito D. et al., 2009; Di Caro D. et al., J Mol Biol 2004; Cavalieri V. et al., NAR 2009). In this study we extend these findings, demonstrating that sns5 works as chromatin insulator also when placed in flanking position of a GFP transgene contained in a lentivirus vector (LV-GFP). A large panel of mouse erythroleukemic clones (MELC) was generated after transduction with uninsulated and sns5 -insulated LV-GFP. Individual clones were screened for single vector integrants (by Q-PCR), and for GFP-expression (by cytofluorimetry). Our results shown that the inclusion of the sns5 element in a forward orientation increased the fraction of vector expressing cells (89% for the insulated vector vs 42% for the uninsulated ones). The clonal variegation of expression, assessed as frequency of clones that showed a percentage of GFP-negative cells in the progeny, decreased in clones transduced with the insulated vectors (7.4% vs 13,9%). It has been suggested that chromatin insulators could shape the architecture of topologically independent chromosome domains. High resolution mapping of chromosomal domains in drosophila and higher eukaryotes highlighted that chromatin insulators play a critical role in shaping the architectural genome organization both in a local chromosome environment and in long range chromosomal interaction. Intriguingly, by using the Chromosome Conformation Capture (3C) technology, we demonstrated that the sns5 -flanked LV-GFP integrated at a single copy in the erythroid cell genome is organized into an independent chromatin loop at the integration site. Worth to mention, no looping was detected in the absence of sns5, indicating that the two flanking copies of sns5 are specifically involved in the reorganization of the chromatin structure at the provirus locus. In conclusion our results not only confirm the conserved and striking boundary function of sns5, but also provide a new clue concerning the molecular mechanism that allows this function to occur. On these basis, our findings reassure the use of sns5 to improve both efficacy and safety of lentiviral vectors for gene therapy. This work was funded by the Assessorato Regionale della Salute, Regione Siciliana (PO FESR 4.1.1.1 RIMEDRI) Disclosures No relevant conflicts of interest to declare.

Il presente lavoro di tesi ha lo scopo di comprendere come l'ambiente sociale e le interazioni interpersonali influenzino la salute mentale e il benessere degli individui gender-variant, affrontando alcune delle questioni rilevanti nel campo dal punto di vista della psicologia sociale e clinica. I primi tre studi presentati nella tesi adottano la prospettiva della psicologia sociale per studiare le esperienze di discriminazione affrontate dagli individui transgender in contesti di salute mentale. Esamineremo il problema da due diversi punti di vista: quello degli psicoterapeuti e quello dei clienti transgender che si sono avvicinati ai servizi di salute mentale. Il primo studio mira a studiare il ruolo del pregiudizio anti-transgender nella valutazione psicologica dei pazienti transgender (vs. cisgender) in un campione di campione di psicoterapeute esperte. Il secondo studio è volto a indagare un campione di psicoterapeuti sia microaggressivo nei confronti di clienti lesbiche o transgender (rispetto a una cliente eterosessuale cisgender). Il terzo studio assume la prospettiva delle persone transgender che accedono ai contesti di salute mentale. Al fine di fornire una maggiore comprensione delle esperienze delle persone transgender che accedono a un servizio di psicoterapia, lo studio si propone di indagare le loro esperienze positive di microaffermazione dell’identità all'interno di una relazione terapeutica. Gli ultimi due studi saranno focalizzati sulle conseguenze del minority stress per la salute mentale e il benessere degli individui transgender. Il quarto studio è intrinsecamente clinico e si focalizzerà sull'indagine della personalità di uomini e donne transgender medicalizzati, valutando sia i domini di personalità dimensionali proposti dal Modello Alternativo di Disturbi della personalità sia le diagnosi categoriche del Disturbo di personalità DSM-IV . Il quinto studio esaminerà gli effetti delle norme sociali di genere in un diverso dominio psicologico, cioè la sessualità. In particolare, lo studio si concentrerà sull'immaginario e sulla fantasia sessuale, che sono aspetti chiave della sessualità umana in quanto associati all'eccitazione e alla risposta sessuale.
The present work is aimed at understanding how social environment and interpersonal interactions influence the mental health and well-being of gender-diverse individuals, approaching some of the relevant issues in the field from the perspective of both social and clinical psychology. The first three studies presented in the thesis adopt the perspective of social psychology to investigate the experiences of discrimination faced by transgender individuals in mental health contexts. We will look at the problem from two different viewpoint: the one of psychotherapists and the one of transgender clients that approached mental health services. The first study aims at investigating the role of anti-transgender bias in the psychological assessment of transgender (vs. cisgender) patients in a sample of female sample of licensed psychotherapists. The second study aims at investigating whether microaggressions are perpetrated by psychotherapists when confronted to lesbian or transgender fictitious clients (vs. cisgender heterosexual). The third study assumes the perspective of transgender people accessing mental health contexts. In order to provide an understanding of transgender peoples’ help-seeking experiences, the study aimed to investigate their positive experiences of identity microaffirmations within a therapeutic relationship. The last two studies will be focused on the consequences of minority stress for the mental health and well-being of transgender individuals. The fourth study is inherently clinical and will be focused on the investigation of personality patterns of medicalized transgender men and women, by evaluating both the dimensional personality domains proposed by the Alternative Model of Personality Disorders and the categorical DSM-IV personality disorder (PD) diagnoses. The fifth study will look at the effects of societal gender norms in a different psychological domain, that is sexuality. In particular, the study will focus on sexual

Oncolytic virus (OV)-based therapies have an emerging role in the treatment of solid tumors, involving both direct cell lysis and immunogenic cell death. Nonetheless, tumor- associated stroma limits the efficacy of oncolytic viruses by forming a barrier that block efficient viral penetration and spread. Another important hurdle for the efficacy of OVs is the antiviral immune responses, where virus-specific infiltrating T cells clear adenovirus-infected cells without compromising tumor burden. In this thesis, these hurdles have been addressed in separate chapters. We first hypothesized that arming an oncolytic adenovirus with a FAP-targeting bispecific T cell engager (FBiTE) could retarget infiltrated lymphocytes towards cancer-associated fibroblasts (CAFs), enhancing viral spread and favoring antitumor rather than anti-viral immune responses. The engineered ICO15K-expressing FBiTE virus showed similar infectivity and replication potency than the non-armed virus. FBiTE-mediated binding of CD3+ effector T cells and FAP+ target cells led to T-cell activation, proliferation, and cytotoxicity against FAP-positive cells in vitro. In vivo, FBiTE expression increased intratumoral accumulation of T cells and decreased the level of FAP, a marker of CAFs, in tumors. Finally, the antitumor activity of the FBiTE-armed adenovirus was superior to the parental virus. The data presented in this thesis strongly supports that the combination of viral oncolysis of cancer cells and FBiTE-mediated cytotoxicity of FAP- expressing CAFs might be an effective strategy to overcome a key limitation of oncolytic virotherapy, encouraging its further clinical development. Aiming to induce stroma disruption, we secondly generated a panel of oncolytic adenoviruses expressing FAP-targeting immunotoxins and a nitroreductase (NfrA)- activatable prodrug. During the development of these projects, we successfully rescued and characterized all the viruses. However, although immunotoxins molecules were properly expressed and secreted from modified-virus infected cells, no promising results were obtained. In contrast, NfrA-armed virus showed replication-dependent enzymatic activity on target cells, leading to increased oncolytic potency in vitro. These preliminary results indicate that this last strategy could be considered to foster viral spread in stroma-abundant tumors, encouraging its validation in an in vivo setting.
Les teràpies basades en virus oncolítics pel tractament de tumors sòlids es consideren molt prometedores degut a la seva capacitat de combinar la lisi directa de cèl·lules canceroses i la mort cel·lular per l’activació del sistema immune. No obstant, l’estroma associat al càncer forma una barrera que bloqueja la penetració i distribució del virus en el tumor, limitant l’eficàcia dels virus oncolítics. Una altra limitació important és la resposta immune contra el virus. Les cèl·lules T citotòxiques específiques contra el virus que infiltren el tumor eliminen, normalment, les cèl·lules infectades per l’adenovirus sense comprometre la massa tumoral. En aquesta tesi, aquestes limitacions han estat abordades en capítols separats. Primer vam hipotetitzar que un adenovirus oncolític armat amb un bispecific T cell engager (BiTE) contra FAP (FBiTE) podria redirigir els limfòcits infiltrats contra els fibroblasts associats al càncer (CAFs), millorant la distribució viral i afavorint la resposta antitumoral vers l’antiviral. El virus ICO15K que expressa el FBiTE va mostrar un patró d’infectivitat i de replicació similars al virus no armat. La unió de les cèl·lules T efectores CD3+ i les cèl·lules diana FAP+ mitjançada pel FBiTE va provocar l’activació, la proliferació i la citotoxicitat de les cèl·lules T contra la cèl·lules FAP positives in vitro. In vivo, l’expressió de FBiTE va induir l’acumulació intratumoral de les cèl·lules T i la disminució dels nivells de FAP, un marcador de CAFs, en els tumors. Finalment, l’activitat antitumoral dels adenovirus armats amb el FBiTE va ser superior que la del virus parental. Els resultats presentats en aquesta tesi aporten fortes evidències que la combinació de l’oncolisi viral de les cèl·lules canceroses i la citotoxicitat dels CAFs FAP+ mitjançada pel FBiTE pot ser una estratègia efectiva per superar les limitacions claus de la viroteràpia. Aquests resultats incentiven el desenvolupament d’aquesta estratègia pel seu ús en la clínica. Amb l’objectiu de destruir l’estroma, vam generar un panell d’adenovirus oncolítics que expressaven diferents immunotoxines específiques contra FAP i una nitroreducatasa (NfrA) activadora de prodroga. Durant el desenvolupament d’aquests projectes, vam obtenir i caracteritzar tots els virus. No obstant, encara que les diferents immunotoxines van ser adequadament expressades i secretades per les cèl·lules infectades pels virus, no vam obtenir cap resultat prometedor. El virus armat amb la NfrA, en canvi, va mostrar una activació enzimàtica depenent de la replicació del virus en les cèl·lules diana, incrementant la potència oncolítica del virus in vitro. Aquests resultats preliminars indiquen que aquesta última estratègia podria fomentar la distribució viral en tumors rics en estroma i incentiven la seva validació en models animals.

Oncolytic virotherapy with Adenoviruses has regained importance in the past years with the appearance of fresh and promising strategies to deal with tumors. Among the major limitations of this therapy are the immune suppression induced in the tumor microenvironment, which prevents the generation of an antitumor immune response, and the presence of stromal barriers which hinder the viral spread and also contain fibroblasts, cells which are highly resistant to viral replication. In this thesis, both limitations have been addressed in separate chapters. Firstly, aiming to induce immune activation, a panel of viruses expressing, CD200, an immune checkpoint, CD200tr, an N-terminal truncated version of the former with antagonistic effect on its receptor (CD200R), K14, a CD200 homolog from HHV-8, and K14tr, a truncated version of K14 and hence a putative antagonist to CD200R, was generated. Throughout the development of this project, we validated the viability of these viruses, we detected the transgenes in supernatants from infected cultures, we confirmed the inhibitory role of CD200 and K14 and the antagonistic one for CD200tr, but our data did not suggest a similar function for K14tr. As for the second project, we generated oncolytic Adenoviruses expressing bacteria- derived toxins modified in such a way they become activated only stroma-specific proteases, aiming to induce indiscriminate cell death once activated at the target tissue. Alpha-toxin from Clostridium septicum and aerolysin from Aeromonas hydrophyla were the toxins of choice. During the development of this project, we successfully generated and characterized all the viruses, we detected aerolysin in supernatants from infected cultures, we confirmed toxin-mediated cytotoxicity in cultures that expressed the activating proteases, and we performed in vivo studies to evaluate the antitumor efficacy, toxicity and the effects on the stroma of the toxins. Whilst for Alpha-toxin no promising results were obtained, the aerolysin-expressing virus increased oncolytic potency in our models, indicating that it could be considered as a potential clinical candidate in stroma-abundant tumors and encouraging to follow this research pipeline.
La viroteràpia amb Adenovirus oncolítics ha recuperat una embranzida que havia perdut fa anys amb l’aparició de noves estratègies per atacar els tumors. Entre les limitacions més importants que troba aquest tipus de teràpia es troben la immunosupressió induïda en el microambient tumoral, que evita la generació d’una resposta immune antitumoral, i la presència de barreres estromals, que dificulten la dispersió del virus dins el tumor i que conté fibroblasts, cèl·lules molt resistents a la replicació viral. En aquesta tesi s’han adreçat aquests dos problemes en dos capítols diferents. En primer lloc, amb l’objectiu de trobar una manera d’activar les cèl·lules del sistema immune, es va generar una bateria de virus expressant versions solubles de la proteïna humana CD200, un ligand immunoinhibidor; CD200tr, una versió truncada en un domini N- terminal de la primera que té una funció antagonista amb el seu receptor; K14, una proteïna del HHV-8 amb estructura i funció homòlogues a CD200; i K14tr, una versió truncada de K14 que es va testar com a possible antagonista alternatiu a CD200R, el receptor de CD200. En el desenvolupament d’aquest projecte, es va validar la viabilitat d’aquests virus, es va detectar transgen en sobrenedants de cultius infectats, es va confirmar el paper inhibidor de CD200 i K14 i l’antagonista de CD200tr, però no es van trobar indicis que K14tr pogués actuar de la mateixa manera. Quant a al segon projecte, es van generar virus oncolítics expressant toxines bacterianes modificades per activar-se tan sols en presència de proteases específiques de l’estroma tumoral, amb l’objectiu d’induir una mort cel·lular indiscriminada un cop activades al teixit diana. Les toxines escollides van ser l’Alpha-toxin de Clostridium septicum i l’aerolisina d’Aeromonas hydrophyla. Durant el desenvolupament del projecte es van generar i caracteritzar satisfactòriament tots els virus, es va detectar transgen en sobrenedants de cultius infectats, es va confirmar l’activitat citotòxica d’aquestes toxines en cèl·lules que expressaven les proteases a les quals havien estat dirigides, i es van fer estudis in vivo per avaluar l’eficàcia antitumoral, la toxicitat i l’efecte en l’estructura de l’estroma de les esmentades toxines. Mentre que l’Alpha-toxin no va generar resultats prometedors, els resultats obtinguts amb el virus expressant aerolisina obren la porta a considerar-lo com a un candidat clínic en tumors amb alt contingut estromal i a seguir aquesta línia de recerca.

La viroterapia oncolítica es un tratamiento emergente basado en el uso de virus replicativos que de manera selectiva infectan células tumorales y provocan su muerte. Hasta la fecha, se han producido grandes avances en dotar a los virus de selectividad tumoral. Sin embargo, el principal desafío actual es potenciar la eficacia de este abordaje a fin de poder implementarlo en la práctica clínica habitual. Una de las estrategias más prometedoras para dotar de mayor potencia antitumoral a los virus oncolíticos, pasa por incorporar genes terapéuticos en el genoma viral, a fin de que se expresen en el microambiente tumoral y aporten beneficios adicionales a la oncolisis. No obstante, se ha observado que, en ocasiones, la expresión de una proteína terapéutica puede afectar negativamente al fitness viral, haciendo que la capacidad replicativa del virus disminuya de manera considerable. Estudios previos indican que el adenovirus utiliza la desviación en su uso de codones como mecanismo para optimizar el reparto de los recursos traduccionales de la célula. En base a este hecho, hemos hipotetizado que la expresión de transgenes terapéuticos podría afectar al equilibrio en el 5 uso de codones del genoma adenoviral, y conducir a un fenómeno de competición intergénica por dichos recursos. A fin de determinar el impacto del uso de codones de los transgenes armados en el genoma adenoviral, hemos estudiado cómo un mismo transgén con distintos patrones de uso de codones afecta a la expresión de las proteínas virales y a la producción de nuevos viriones. Nuestros datos sugieren que los transgenes, cuando se expresan en la fase tardía pos- infección, compiten con los genes virales por los recursos traduccionales, afectando el fitness viral de manera dependiente al uso de codones. Transgenes altamente optimizados, secuestrarían los recursos traduccionales de la célula en detrimento de los genes virales, conduciendo así a una replicación viral deficiente. No obstante, hemos demostrado que es posible rescatar la actividad viral mediante la modulación del grado de optimización de los transgenes. Estos resultados se han evidenciado en adenovirus armados con genes codificantes para la proteína reportera verde fluorescente o GFP, así como con dos transgenes terapéuticos, la enzima hialuronidasa (que degrada la matriz celular y reduce la desmoplasia tumoral) y la enzima LmPDT (que cataliza la conversión de prodrogas en bases púricas citotóxicas). En conjunto, concluimos que adaptar el uso de codones de los transgenes armados en el genoma adenoviral es un parámetro crítico que debería considerarse en el diseño de adenovirus oncolíticos armados a fin maximizar el beneficio terapéutico de los mismos.

Therapeutic angiogenesis is the attempt to increase vascular density by means of an exogenously administered proangiogenic agent and offers a potential treatment for diseases associated with tissue ischemia. Vascular endothelial growth factor (VEGF) expressed by gene therapy has been shown to be a potent stimulator of angiogenesis and to improve the function of ischemic tissues in patients [Isner, 1998]. Unregulated gene therapy is disconcerting since there is no assurance that the treatment will target the ischemic territory. A new regulated adeno-associated viral vector expressing VEGF165 that is conditionally silenced has been developed by one of the authors (KAW). The transgene expression is regulated by silencing the genes in the absence of the disease and at the same time having strong and local activation in the presence of the disease. The purpose of this work is to establish protocols and techniques to quantify the efficacy of therapeutic angiogenesis. The initial phase of this research involves assessment of angiogenesis using an unregulated, adenoviral vector that is encoded to express VEGF165. Using the rabbit hind limb ischemia model, angiography was performed on animals that were given the proangiogenic treatment and on a sham group, in which phosphate buffered saline (PBS) was injected. Angiographic contrast intensity curves were obtained, modeled, and the optimized model parameters provided insight into flow characteristics within the targeted vascular bed. In the second phase of the project the conditionally silent vector will be employed using the developed protocols and methods of the first phase to afford comparisons with the previous groups.