La Genetica del Giudizio Morale: le scelte di natura morale possono essere influenzate dal nostro patrimonio genetico?

Riassunto La comprensione dei meccanismi neurobiologici che sottendono il comportamento sociale umano è diventata, negli ultimi anni, oggetto di numerose ricerche. E’ stato osservato, ad esempio, che molti tratti sociali, inclusi quelli che influenzano le scelte morali quali l’empatia, l’altruismo, l’impulsività e la capacità di reagire allo stress, mostrano un’ereditarietà compresa tra il 10% e il 60%. Non è noto, però, ad oggi, quali varianti genetiche giochino un ruolo nei meccanismi decisionali alla base del giudizio morale, per cui la loro identificazione appare di grande interesse. In questo lavoro di tesi abbiamo deciso di indagare se alcuni polimorfismi genetici associati al comportamento impulsivo possono influenzare le scelte morali. L'impulsività, infatti, è un tratto di personalità che, pur associandosi talvolta a comportamenti patologici, consente all’individuo di prendere decisioni tempestive, in situazioni che lo richiedono, influenzando pertanto anche le scelte morali. Poiché uno dei meccanismi alla base del comportamento impulsivo sembra essere l’alterazione dei livelli cerebrali di dopamina, sono state selezionate 4 varianti localizzate su 3 geni implicati nella regolazione del sistema dopaminergico: la VNTR (Variable Number of Tandem Repeats) SLC6A3–3/11 presente nel 3’UTR (UnTranslated Region) del gene codificante per il trasportatore della dopamina (SLC6A3), la VNTR DRD4–2/11 presente nell’esone 3 del gene codificante per il recettore dopaminergico D4 (DRD4), lo SNP rs1800955 presente nel promotore del gene DRD4 e lo SNP rs4680 presente nell’esone 4 del gene codificante per la Catecol-O-metiltransferasi (COMT), che è un enzima deputato al metabolismo dei neurotrasmettitori, compresa la dopamina. Il DNA è stato estratto dalla saliva di 200 soggetti sani a cui è stato chiesto di rispondere a una serie di dilemmi morali per valutare la loro tendenza ad agire. Data la differente tipologia dei polimorfismi selezionati, tre tecniche di biologia molecolare sono state utilizzate per la loro genotipizzazione: l’High Resolution Melting (HRM)-PCR per lo SNP rs4680, la PCR-RFLP (Restriction Fragment Lenght Polymorphism) per gli SNP rs1800955 e la PCR con successiva corsa elettroforetica su gel di agarosio al 2% per l’analisi delle VNTR. I casi dubbi sono stati risolti mediante sequenziamento di Sanger. I nostri dati hanno mostrato che, nonostante le decisioni morali siano influenzate per lo più dalla tipologia di dilemma, le risposte dei soggetti sono comunque condizionate dal loro genotipo. In particolare, lo SNP rs4680 sul gene COMT si è dimostrato come quello maggiormente associato alle risposte affermative. Gli altri polimorfismi mostrano associazione soltanto se in interazione tra di loro o con lo scenario proposto nel dilemma. Abstract The comprehension of the neurobiological mechanisms underlying human social behavior has become, in recent years, the subject of numerous researches. Most human behavioral/social traits, such as altruism, empathy and impulsivity, show high hereditability, indicating that the genetic background exerts a considerable influence on behavior. It is still unknown, however, which genetic variants play a role in the decision-making mechanisms at the basis of moral judgment, whereby their identification appears of great interest. The aim of this thesis was to investigate whether certain genetic polymorphisms associated with impulsive behavior could influence moral choices. Impulsivity is, in fact, a personality trait that, although sometimes is associated with pathological behaviors, enables the individual to make timely decisions in situations that require it, therefore affecting moral choices. Since one of the mechanisms underlying impulsive behavior seems to be the alteration of brain levels of dopamine, we selected five variants localized on four genes involved in the regulation of the dopaminergic system: the VNTR (Variable Number of Tandem Repeats) SLC6A3–3/11 located in the 3'UTR (untranslated Region) of the gene encoding the dopamine transporter (SLC6A3), the VNTR DRD4-2/11 located in exon 3 of the gene encoding the dopamine receptor D4 (DRD4), the SNP rs1800955 located in the promoter of the DRD4 gene and the SNP rs4680 located in exon 4 of the gene encoding the Catechol-O-methyltransferase (COMT), which is an enzyme responsible for the metabolism of neurotransmitters, including dopamine. From an experimental point of view, DNA was collected by taking saliva samples from 200 healthy volunteers, who were asked to answer a series of moral dilemmas proposed by a team of psychologists, in order to assess their tendency to act, the response time, the emotional involvement and the moral acceptability of the newly accomplished. My experimental thesis work consisted in genotyping 200 samples for the selected genetic variants. Three molecular biology techniques have been used for their genotyping: the High Resolution Melting (HRM)-PCR for the SNP rs4680, the PCR-RFLP (Restriction Fragment Lenght Polymorphism) for the SNP rs1800955 and PCR (Polimerase Chain Reaction) followed by electrophoresis on agarose gel for the VNTR analysis. The doubtful cases were solved by Sanger’s sequencing. Our results provide evidence that, despite the moral decisions are influenced mostly by the type of dilemma, the subjects answers are conditioned by their genotype. In particular, the SNP COMT rs4680 showed the highest association with the affirmative responses. The other polymorphisms showed association only when in interaction to each other or to the scenario proposed by the dilemmas

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

Characterization of breast cancer susceptibility genes in men by Next-Generation Sequencing

Male breast cancer (MBC) is a rare disease with an incidence of 1/105 year. MBC tends to occur between 60-70 years old and to express oestrogen and progesterone receptors with occasionally epidermal growth factor receptor 2 amplification. The rarity of MBC has precluded large-scale association studies, thus genetic predisposition remain not well understood. In order to better define genetic risk factors in men, a germline investigation in 81 MBC cases was performed through the screening of 24 genes involved in breast cancer predisposition, genome stability maintenance and DNA repair mechanisms by Next Generation Sequencing. Clinical pathological data and family history of 81 MBC cases were collection revealed an average age of onset was 61.3 years and breast cancer family history in 35 cases. Overall, our genetic screening let to attribute a genetic cause to breast cancer in 23% of cases. In total, 19 patients carried a pathogenic mutation in 4 genes: BRCA2, BRIP1, MUTYH and PMS2. As expected, a positive family history is a strong predictor of germline BRCA2 mutations. Moreover, 14 variants of unknown clinical significance (VUS) in 9 genes (BARD1, BRCA1, BRCA2, BRIP1, CHEK2, ERCC1, NBN, PALB2, PMS1) were predicted as potentially pathogenic by in-silico analysis leading to 40% the mutation detection rate. Understanding the potential pathogenicity of VUS represents an extremely urgent question for the management of breast cancer risk in MBC cases and their own families

Is it time to consider the Androgen receptor as a therapeutic target in breast cancer?

: Breast cancer (BC) is a heterogeneous disease and the most prevalent malignant tumor in women worldwide. The majority of BC cases are positive for estrogen receptor (ER) and progesterone receptor (PgR), both known to be involved in cancer pathogenesis, progression, and invasion. In line with this, hormonal deprivation therapy appears to be a useful tool and an effective treatment for these BC subtypes. Unfortunately, prognosis among patients with hormone-negative tumors or therapy-refractory and metastatic patients remains poor. Novel biomarkers are urgently needed in order to predict the course of the disease, make better therapy decisions and improve the overall survival of patients. In this respect, the androgen receptor (AR), a member of the hormonal nuclear receptor superfamily and ER and PgR, emerges as an interesting feature widely expressed in human BCs. Despite the advances, the precise tumorigenic mechanism of AR and the role of its endogenous ligands are yet not well-understood. In this review, we aim to elaborate on the prognostic impact of AR expression and current AR-targeting approaches based on previous studies investigating AR's role in different BC subtypes

Androgen receptor expression inversely correlates with histological grade and N stage in ER+/PgRlow male breast cancer

Androgen Receptor (AR) positivity is often displayed in breast cancer and especially in Male Breast Cancer (MBC), where it appears to be a heterogeneous feature, with its expression ranging between 38 and 81% of cases. Given the fact that circulating androgens represent the most important sex hormones in males and that breast carcinogenesis is characteristically subjected to hormonal mechanisms, our purpose was to investigate the clinicopathological significance of AR in MBC assessing if its expression could be associated with parameters of tumor aggressiveness

Diagnosis and treatment monitoring in breast cancer: how liquid biopsy can support patient management

Imaging and tissue biopsies represent the current gold standard for breast cancer diagnosis and patient management. However, these practices are time-consuming, expensive and require invasive procedures. Moreover, tissue biopsies do not capture spatial and temporal tumor heterogeneity. Conversely, liquid biopsy, which includes circulating tumor cells, circulating free nucleic acids and extracellular vesicles, is minimally invasive, easy to perform and can be repeated during a patient's follow-up. Increasing evidence also suggests that liquid biopsy can be used to efficiently screen and diagnose tumors at an early stage, and to monitor changes in the tumor molecular profile. In the present review, clinical applications and prospects are discussed

The Prognostic Impact of Gender, Therapeutic Strategies, Molecular Background, and Tumor-Infiltrating Lymphocytes in Glioblastoma: A Still Unsolved Jigsaw

Despite the adoption of novel therapeutical approaches, the outcomes for glioblastoma (GBM) patients remain poor. In the present study, we investigated the prognostic impact of several clinico-pathological and molecular features as well as the role of the cellular immune response in a series of 59 GBM. CD4+ and CD8+ tumor-infiltrating lymphocytes (TILs) were digitally assessed on tissue microarray cores and their prognostic role was investigated. Moreover, the impact of other clinico-pathological features was evaluated. The number of CD4+ and CD8+ is higher in GBM tissue compared to normal brain tissue (p p = 0.0005 respectively). A positive correlation between CD4+ and CD8+ in GBM is present (rs = 0.417—p = 0.001). CD4+ TILs are inversely related to overall survival (OS) (HR = 1.79, 95% CI 1.1–3.1, p = 0.035). The presence of low CD4+ TILs combined with low CD8+ TILs is an independent predictor of longer OS (HR 0.38, 95% CI 0.18–0.79, p = 0.014). Female sex is independently related to longer OS (HR 0.42, 95% CI 0.22–0.77, p = 0.006). Adjuvant treatment, methylguanine methyltransferase (MGMT) promoter methylation, and age remain important prognostic factors but are influenced by other features. Adaptive cell-mediated immunity can affect the outcomes of GBM patients. Further studies are needed to elucidate the commitment of the CD4+ cells and the effects of different TILs subpopulations in GBM

Forecasting Molecular Features in IDH-Wildtype Gliomas: The State of the Art of Radiomics Applied to Neurosurgery

Background: The fifth edition of the WHO Classification of Tumors of the Central Nervous System (CNS), published in 2021, marks a step forward the future diagnostic approach to these neoplasms. Alongside this, radiomics has experienced rapid evolution over the last several years, allowing us to correlate tumor imaging heterogeneity with a wide range of tumor molecular and subcellular features. Radiomics is a translational field focused on decoding conventional imaging data to extrapolate the molecular and prognostic features of tumors such as gliomas. We herein analyze the state-of-the-art of radiomics applied to glioblastoma, with the goal to estimate its current clinical impact and potential perspectives in relation to well-rounded patient management, including the end-of-life stage. Methods: A literature review was performed on the PubMed, MEDLINE and Scopus databases using the following search items: “radiomics and glioma”, “radiomics and glioblastoma”, “radiomics and glioma and IDH”, “radiomics and glioma and TERT promoter”, “radiomics and glioma and EGFR”, “radiomics and glioma and chromosome”. Results: A total of 719 articles were screened. Further quantitative and qualitative analysis allowed us to finally include 11 papers. This analysis shows that radiomics is rapidly evolving towards a reliable tool. Conclusions: Further studies are necessary to adjust radiomics’ potential to the newest molecular requirements pointed out by the 2021 WHO classification of CNS tumors. At a glance, its application in the clinical routine could be beneficial to achieve a timely diagnosis, especially for those patients not eligible for surgery and/or adjuvant therapies but still deserving palliative and supportive care

Forecasting Molecular Features in IDH-Wildtype Gliomas: The State of the Art of Radiomics Applied to Neurosurgery

Background: The fifth edition of the WHO Classification of Tumors of the Central Nervous System (CNS), published in 2021, marks a step forward the future diagnostic approach to these neoplasms. Alongside this, radiomics has experienced rapid evolution over the last several years, allowing us to correlate tumor imaging heterogeneity with a wide range of tumor molecular and subcellular features. Radiomics is a translational field focused on decoding conventional imaging data to extrapolate the molecular and prognostic features of tumors such as gliomas. We herein analyze the state-of-the-art of radiomics applied to glioblastoma, with the goal to estimate its current clinical impact and potential perspectives in relation to well-rounded patient management, including the end-of-life stage. Methods: A literature review was performed on the PubMed, MEDLINE and Scopus databases using the following search items: “radiomics and glioma”, “radiomics and glioblastoma”, “radiomics and glioma and IDH”, “radiomics and glioma and TERT promoter”, “radiomics and glioma and EGFR”, “radiomics and glioma and chromosome”. Results: A total of 719 articles were screened. Further quantitative and qualitative analysis allowed us to finally include 11 papers. This analysis shows that radiomics is rapidly evolving towards a reliable tool. Conclusions: Further studies are necessary to adjust radiomics’ potential to the newest molecular requirements pointed out by the 2021 WHO classification of CNS tumors. At a glance, its application in the clinical routine could be beneficial to achieve a timely diagnosis, especially for those patients not eligible for surgery and/or adjuvant therapies but still deserving palliative and supportive care