FIRST YEAR REPORTS
The main lines of research concerned:
Evaluation of ovarian steroidogenesis in patients with endometriosis
Endometriosis is an estrogen-dependent pelvic inflammatory disease characterized by the implantation and growth of endometrial tissue (glands and stroma) outside the uterine cavity. It may be asymptomatic or associated with pain symptoms (chronic pelvic pain, dysmenorrhea and dyspareunia) and/or infertility. The association between endometriosis and infertility is well known. Although the literature extensively addresses the association between the disease and infertility, the etiopathogenetic mechanisms involved in this relationship are not yet fully understood. Among the various potentially harmful mechanisms there is also the possibility of an intraovarian endocrine deregulation. Inadequate steroid hormone production could be implicated in impaired follicular function, a situation that could negatively affect oocyte quality. Currently, the evidence regarding the correlation between the levels of hormones in the follicular fluid (FF) and the outcomes of the PMA cycle in patients with endometriosis is, however, partial and incomplete.
The primary objective of the research project is to compare the levels of 15 steroid hormones in the follicular fluid of infertile women with and without endometriosis undergoing in vitro fertilization (IVF).
The secondary objectives are as follows:
1) Correlate intraovarian hormone levels with clinical and embryological outcomes (diameter and number of follicles, number of oocytes retrieved, number of mature oocytes, number of grade I and II embryos formed and pregnancy rate).
2) Compare markers of ovarian reserve, expressed as anti-Müllerian hormone (AMH), antral follicle count (AFC), and age, with intraovarian hormone levels.
3) Evaluate the independent effect on steroidogenesis of the presence of ovarian endometriotic cysts.
4) Analyze the levels of endocrine pollutants in the follicular fluid (phthalates) and correlate them with intraovarian steroid hormone levels and the presence/absence of endometriosis.
Patients affected by endometriosis (ultrasound or histological diagnosis) and candidates for second level PMA techniques will be included; controls will match the next patient free of disease.
This single-centre prospective cohort study envisages the recruitment of patients with endometriosis who will undergo second-level PMA treatments during the observation period. For each patient with endometriosis, the next infertile patient due to other causes (tubal factor, ovulatory disorders, reduced ovarian reserve, male infertility and unexplained infertility) will be matched.
As per the operating procedure, the follicular fluid aspirated during the oocyte retrieval procedure will be delivered to the biologists so that they can identify and isolate the oocytes. In this study, the follicular fluid, which is usually eliminated, will be collected in test tubes. All samples will be thawed simultaneously at the time of analysis (steroidogenic cascade and phthalates). For women with unilateral endometriomas, you will get two separate pools for the two ovaries.
Following the clinical practice, the patients enrolled in the study will continue the regular path of assisted reproduction.
Data will be analyzed using the Statistical Package for Social Sciences (SPSS 26.0, IL, USA). Student's t-test or Mann-Whitney nonparametric test will be used for quantitative variables and Pearson's x2 test or Fisher's exact test for qualitative variables, as appropriate. The evaluation of the associations between phthalates and steroid hormones will be explored by means of correlation tests (Spearmann, given the non-normality of the variables). Considering that non-parametric tests will be used, a sample size of 160 women (80 cases and 80 controls) will allow, by applying the usual type I and II errors of 0.05 and 0.20, to highlight differences such that, in patients with endometriosis, 25% will have values higher than the 90th centile of the distribution of values in the controls (Odd ratio of values higher than the 90th centile ≥ 3).
Diabetes Mellitus type 2 and Non-Alcoholic Fatty Liver Disease: A complete evaluation in terms of prevalence, risk factors and clinical relevance.
During the last decades, there have been changes in people's lifestyles with significant consequences on human metabolism. Therefore, chronic metabolic diseases such as obesity, type 2 diabetes mellitus (type 2 DM), and nonalcoholic fatty liver disease (NAFLD) have exploded worldwide. Scientific evidence has pointed out a significant association of type 2 DM and NAFLD. In fact, diabetes, despite being a risk factor for the development of NAFLD, also appears to accelerate its progression. Different studies have investigated the prevalence of NAFLD in type 2 DM; however, these results are inconsistent.
1) Assessment of the prevalence of NAFLD inT2DM patients; 2) evaluation of clinical presentation of NAFLD inT2DM patients; 3) identification of the general, clinical and biochemical characteristics of T2DM patients with and without NAFLD; 4) evaluation of the association of clinical and biochemical characteristics in patients with type 2 diabetes with and without NAFLD; 5) investigation of vascular adhesion molecule -1 (VCAM-1), intercellular adhesion molecule -1 (ICAM-1), E-selectin levels in the sera of patients with T2DM with and without NAFLD.
80 patients with T2DM were included in this study. Each patient's anamnestic, clinical and laboratory data were registered. The presence of NAFLD was assessed using ultrasound. Commercial ELISA kits were used to detect the presence of VCAM-1, ICAM-1, E-selectin in the serum of patients with type 2 DM with and without NAFLD. Statistical analysis helped us to study the significant relationships of the evaluated parameters in T2DM patients with and without NAFLD.
80 patients diagnosed with T2DM for an average of 10.1 years (47F/33M) with a mean age of 60.4 years were included in this study. Hypertension (76.2%), Dyslipidemia (82.5%) and Central Obesity (81.3%) were the most common comorbidities. The prevalence of NAFLD in patients with T2DM was 61.3% and the following grades of NAFLD were evidenced by ultrasound: 31 (38.8%) patients grade 0; 19 (23.7%) grade 1 patients; 22 (27.5%) grade 2 patients; 8 (10%) grade 3 patients. Of 49 T2DM patients also diagnosed with NAFLD, 37 (76%) patients were asymptomatic, while 12 (24%) patients presented symptoms of NAFLD. The most frequent symptoms were the feeling of fatigue and generalized body weakness, as well as abdominal discomfort mainly localized in the upper right quadrant. The comparison between clinical and laboratory characteristics in patients with T2DM with and without NAFLD resulted in statistically significant differences for the mean values of: BMI (p = 0.001), waist circumference (p = 0.0001), duration in years of DM (p = 0.0001), fasting blood glucose (p = 0.0001), HbA1c (p = 0.0001), insulinemia (p = 0.001), HOMA-IR (p = 0.012), systolic and diastolic BP (p = 0.00006 and p = 0.002), HDL-cholesterol and TG (p = 0.0001 and p = 0.0001), AST (p = 0.006), ALT (p = 0.02), GGT (p = 0.00001), ALP (p = 0.0005). Obesity was the variable with the strongest association with NAFLD in T2DM patients with p = 0.0339, OR = 5.8527 and CI 95% 1.1438 – 29.9484. Serum levels of sICAM-1, sVCAM-1, and sE-selectin were significantly higher in T2DM patients with and without NAFLD compared to healthy controls (1p= 0.0001) while sVCAM-1 levels were significantly higher in the T2DM with NAFLD group than in the T2DM without NAFLD group (2p = 0.001).
NAFLD was present in 61.3% of patients with T2DM, although most of them were asymptomatic (76%). BMI, waist circumference, duration of diabetes in years, fasting blood glucose, HbA1c, insulinemia, HOMA-IR, BP, TG, AST, ALT, GGT and ALP were at higher mean values in patients with type 2 DM and NAFLD; only HDL presents lower mean values in these patients. Obesity was found to be an independent risk factor associated with NAFLD in patients with type 2 DM. Serum levels of sICAM-1, sVCAM-1, and sE-selectin were significantly higher in T2DM patients with and without NAFLD compared to healthy controls, while sVCAM-1 levels were significantly higher in the T2DM with NAFLD group than in the T2DM without NAFLD group. This study confirms that in our diabetic patients, NAFLD is closely related to obesity, insulin resistance and metabolic syndrome, therefore it can also be considered as the hepatic manifestation of metabolic syndrome.
The role of sperm miRNA-34c and miRNA-449b in male infertility
MiR-34c and miR-449b show differential expression in patients with abnormal semen parameters. Furthermore, miR-34c is associated with the outcome of ICSI, in particular with the kinetics of embryonic development and the quality of the embryo. Interest in understanding the role of sperm chromatin integrity has also grown in recent decades. High fragmentation of sperm DNA reduces the fertilization rate and affects the development of the embryo.
In light of this, the aim of this study was to analyze the levels of miR-34c, miR-449 and sperm chromatin integrity in order to identify new biomarkers of semen quality and assisted fertilization outcomes.
The research is divided into two studies. Study 1) analysis of miR-34c levels in spermatozoa of normozoospermic subjects and patients with altered semen characteristics. 20 men belonging to the Andrology clinic of the Department of Experimental Medicine were recruited, of which 10 with alteration of seminal parameters and 10 normozoospermic. These subjects underwent semen examination and molecular analysis of miR-34c and miR-449b. Study 2) analysis of the levels of nemaspermic miR-34c and miR-449 and of the DNA fragmentation of the spermatozoa of men of couples subjected to assisted fertilization. The male partner of 106 couples affected by primary infertility belonging to the PMA Center of the Policlinico Umberto I was recruited. In this group of subjects, semen, ICSI outcomes and sperm DNA fragmentation were evaluated. The expression of miR-34c and miR-449b was evaluated in a subgroup of 38 subjects. For semen analysis, samples were evaluated according to the WHO Laboratory Manual for Semen Examination (2021).
Sperm DNA fragmentation (SDF) was assessed using the TUNEL (Terminal deoxynucleotidyl transferase UTP-driven Nick End Labeling) assay. Profile analysis of miR-34c, miR-499 miRNAs was performed using ddPCR.
Sperm DNA fragmentation is negatively correlated with semen parameters and is reduced after sperm selection. %SDF is also associated with embryo quality and consequently with the number of embryos transferred. In particular, %SDF>2.95% increases the risk of failure of embryonic development by 4 times. miR-34c expression is associated with sperm concentration and motility. miR-449 is associated with concentration and %SDF. Furthermore, the two miRNAs are surprisingly related to each other. An increase of miRNA-34c compared to miR-449b increases the probability of obtaining developing embryos. Conversely, higher levels of miR-449b compared to miR-34c reduces the probability of obtaining developing embryos. These data demonstrate the promising role of miR-34c, miR-449 and %SDF as biomarkers of assisted reproduction outcomes.
Natural inhibitors of phosphodiesterases in the regulation of metabolic alterations induced by exposure to endocrine disruptors
Obesity is an endocrine disease resulting from the interaction between behavioural, social, genetic and environmental factors. Among the environmental factors there is a group of chemical substances, natural and synthetic, known as endocrine disruptors, which act on the endocrine system. There is different experimental evidence on animal models and humans regarding the impact that endocrine disruptors have on metabolic disorders as they are able to reproduce, block or reduce the physiological activity of hormones. The second messenger cGMP is involved in the regulation of energy homeostasis and cellular metabolism. Pharmacological induction of cGMP is known to have beneficial effects in mouse models of the metabolic syndrome. The intracellular levels of cGMP are finely regulated by phosphodiesterases (PDEs) and the main enzyme responsible for its degradation is PDE5. Preclinical and clinical studies have demonstrated that PDE5 inhibitors have beneficial effects on improving insulin resistance and glucose metabolism, representing a promising therapeutic strategy for the treatment of metabolic disorders.
Although several drugs are available to modulate the activity of PDEs, in reality many natural products contain phosphodiesterase inhibitors. Among these, the best known is caffeine, a non-selective inhibitor of phosphodiesterases, which causes an acute increase in the levels of the second messenger cAMP. In reality many other products belong to this class but further studies are needed to investigate the mechanism of action and effectiveness of these compounds.
The aim of this study is to evaluate the role of PDE5 in the metabolic alterations induced by exposure to endocrine disruptors in cell models overexpressing or lacking PDE5 and in a PDE5 knockout mouse model.
- For this purpose, an in vitro cell model was developed using the 293T cell line to determine the concentration and timing of treatment with the endocrine disruptor Bisphenol A (BPA) and to evaluate cell viability.
- A 3T3L1 murine preadipocyte line to evaluate the impact of BPA and the role of PDE5 in modifying the modified metabolic pathways. In particular, I will use systems to modulate PDE5 expression.
- Primary cells isolated from adipose tissue of the PDE5 knockout and wild-type mouse model.
- PDE5 knockout and wild-type mice for acute and chronic BPA treatment cycles
- Natural substances with analogous activity to known natural inhibitors that have an action in mimicking the effects of phosphodiesterase inhibitors on all models used
- Isolated primary cells of adipose tissue to re-screen natural compounds.
New ultrasound parameters predictive of thyroid nodule malignancy
Recently, some new ultrasound methods have been proposed to define the predictive parameters of malignancy of the thyroid nodule:
1) Strain elastography (SE) and, more recently, Shear Wave elastography (SWE). Both offer real-time information on the stiffness of the tissue under examination: the SE provides qualitative and semi-quantitative information and is operator-dependent; the SWE additionally provides quantitative information and is operator-independent. Studies conducted to date on ES have demonstrated high sensitivity and high negative predictive value but low positive predictive value for detecting thyroid carcinomas. SWE has only recently been tested in thyroid nodules.
2) the AngioPL.U.S. (AngioPLane wave Ultra Sensitive): new ultrasensitive Doppler technique that allows to evaluate the microvascular architecture of the nodule and therefore the small blood vessels, where the flow is much slower, typical of tumor neoangiogenesis. To date there are no studies in any application field.
3) TriVu: allows you to view, in "real time" and simultaneously, the B-dimensional image, the elastographic map and the vascularization of the nodule (B-Mode + SWE + Power Doppler). Starting from the assumption that parenchymal areas with intense vascularization have a lower stiffness, thanks to the use of this software, it is possible to sample the tissue stiffness in areas with lower vascular density, thus obtaining a better definition of the stiffness of the extravascular parenchyma. To date, there are still no studies aimed at testing the diagnostic capacity of this software in the field of thyroid nodules.
OBJECTIVE 1: to date 141 nodules have been analyzed to validate the preliminary results obtained from the series of 240 nodules regarding:
- the predictive value of malignancy and the diagnostic accuracy of SWE and AngioPL.U.S.
- the correlation between thyroid nodule volume and SWE and between thyroid nodule volume and AngioPL.U.S.
- the cut-off value of the SWE to discriminate between benign and malignant nodules
- the vascular pattern, using AngioPL.U.S., with better diagnostic accuracy
For these purposes, thyroid ultrasound will be performed in patients with nodular or multinodular thyroid disease, evaluating the "standard" ultrasound parameters for each nodule, the vascularization both by means of conventional Color Doppler and by AngioPL.U.S. and stiffness by SWE (Q-Ratio, Emax, Emin, DS). Each nodule will then be assigned an EU-TIRADS category in accordance with the latest ETA guidelines. Subsequently they will be submitted to FNAC in accordance with the ETA guidelines of 2017 and/or on the basis of the SWE and the AngioPL.U.S. and following surgery (histological examination) on the basis of cytological examination (Tir3b, Tir4, Tir5), risk factors (Tir3a) and the possible presence of compressive symptoms.
OBJECTIVE 2: to date 35 Tir3a nodules have been recruited for:
- evaluate any variations, in terms of "standard" ultrasound parameters, SWE and AngioPL.U.S. and FNAC, in the follow-up of Tir3a. For this purpose, the Tir3a subgroup will undergo follow-up at 3-5 years, with clinical-instrumental checks at 6/12 months based on the risk factors, to evaluate whether the possible appearance of changes in these parameters is associated with changes in the risk of malignancy, confirmed by a new FNAC.
OBJECTIVE 3: to date 14 nodules have been recruited for:
- increase the diagnostic accuracy of FNAC using SWE. For this purpose, an equal number of nodules will be prospectively selected, homogeneous in size (≥ 2 cm) and in ultrasound characteristics (EU-TIRADS 3), which will be divided into 2 groups: group 1 (US-guided FNAC execution using the «conventional method») and group 2 (US-guided FNAC execution using SWE).
- increase the diagnostic accuracy of SWE using TriVu. For this purpose, since the stiffness distribution is not uniform in large nodules and to allow for the exclusion of parenchymal areas with high vascular density and less stiffness, nodules ≥ 2 cm in size will be selected in which both elastography by SW and that via TriVu. The standardization of the execution technique is underway.
MATERIALS
UltraFast ultrasound:
• Supersonic Aixplorer Elite with 15-4 MHz broadband linear probe (50 mm) and 20-6 MHz microlinear probe (27 mm)
• Supersonic Aixplorer Elite MACH30 with 18-5 MHz broadband (50 mm) linear probe
Role of beta arrestins and G proteins in mediating dopamine receptor type 2 (DRD2) signal transduction in pituitary tumors.
Dopamine receptor type 2 (DRD2) agonists are the first-line treatment for prolactin-prolactin (PRL)-producing pituitary tumors, but are poorly effective in non-functioning pituitary neuroendocrine tumors (NF). The reason for the poor responsiveness of NF-PitNETs to DA can be sought in the intracellular events that occur downstream of DRD2 activation. Like many other G protein-coupled receptors, DRD2 mediates both canonical G protein-dependent and non-canonical beta-arrestin-2-dependent signaling pathways. The canonical signaling pathway of DRD2 involves coupling with inhibitory G proteins, inhibition of adenyl cyclase, and subsequent reduction of intracellular cAMP. In parallel, the DRD2/beta-arrestin-2 interaction initiates independent signaling, leading to the reduction of AKT phosphorylation, playing a prominent role for the antiproliferative effect of DRD2 in pituitary tumors. Since functionally selective DRD2 agonists preferentially activate one of these pathways, they represent a promising pharmacological strategy to increase therapeutic efficacy.
The aim of the study program is to investigate the role of G proteins and beta-arrestin 2 in mediating DRD2 signaling in MMQ rat prolactin-secreting tumor cells and in human primary NF-PitNET cells. UNC9994, a selective DRD2 beta-arrestin 2 agonist, and MLS1547, a selective G protein agonist, were used for this purpose.
In the MMQ cell model, we found that treatment with UNC9994 reduced cell proliferation (-41.4 20% at 100 nM, p< 0.001 vs baseline) with greater efficacy than cabergoline (-21 10.9% at 100 nM 10.9%, p<0.001 vs bas), unlike the treatment with MLS1547 which resulted in a slight reduction of cell proliferation (-13.2 7.4% at 100 nM, p<0.05 vs bas). Furthermore, UNC9994 was more effective in reducing AKT phosphorylation (-45.5 16%, p<0.005 vs base) than cabergoline, while no effect on AKT phosphorylation was found after treatment with MLS1547. Consistently, treatments with cabergoline and UNC9994 resulted in a significant reduction of cyclin D3 expression (-13.4 ± 5.4%, p<0.005 vs base and -17.6 ± 7.8%, p<0.005 vs base, respectively), together with a increased expression of p27/Kip1 (+35 ± 20%, p<0.05 vs bas and +41.6 ± 20.5%, p<0.05 vs bas, respectively).
Silencing of beta-arrestin 2 reversed the antiproliferative effects of UNC9994 and cabergoline and their effects on AKT phosphorylation. Pretreatment with pertussis toxin (PTX) maintained the antiproliferative effects of cabergoline (-16.6 ± 3.6%, p<0.001 vs. bas) and UNC9994 (-31 ± 1.9%, p<0.001 vs. bas), while it abolished MLS1547's ability to reduce cell proliferation.
Moreover, in the MMQ cell line, the 24h treatment with MLS1547 results in a marked reduction of PRL secretion (-34.7 ± 29.5% at 100nM µM, p<0.01 vs base), compared to the treatment with UNC9994 (-26.7 ± 15.5% at 100nM µM, p<0.01 vs low) and with cabergoline (-20.3 ± 22.3% at 100nM µM, p<0.01 vs low).
After treatment with MLS1547, cell migration showed a significant reduction (-44 ± 10% at 1 µM, p<0.0005 vs baseline), to a greater extent than cells treated with UNC9994 (-31 ± 19% at 1 µM, p <0.005 vs low) and with cabergoline (-12 ± 23.3% at 1 µM, p<0.05 vs low).
The antiproliferative effects of selective DRD2 agonists were also tested on 17 primary cultures from surgically removed NFPitNETs. 8 of 17 human primary NF-PitNET cell cultures grown in vitro responding to the antiproliferative effects of cabergoline (-26.5 10.8%, p<0.001 vs baseline) showed a significant reduction in cell proliferation after treatments with UNC9994 and MLS1457 (-18.3 11.3%, p<0.0005 vs base, and -10.7 9.3%, p<0.001 vs base, respectively). On the other hand, 4 of the 17 NF-PitNETs that did not respond to cabergoline were also resistant to treatment with UNC9994 and MLS1547.
In conclusion, the data generated so far have demonstrated a relevant role of the beta-arrestin 2-dependent pathway in regulating the inhibitory effects of DRD2 on tumor growth, while canonical G protein-mediated signaling appears to be fundamental in the reduction of PRL secretion. and in the control of cell migration.
Hormone replacement therapy in patients with BRCA1-2 gene mutation undergoing prophylactic adnexectomy
Inactivation of the BRCA1 gene causes 45% of breast cancer susceptibility syndromes and at least 80% of genetic susceptibility syndromes to ovarian and breast cancer. Partial inactivation of the BRCA1 and BRCA2 genes currently represents the main mechanism of familial predisposition to breast and ovarian cancer and is associated with a cumulative risk of the appearance of these neoplasms of over 50%. Several observational studies and meta-analyses have shown that salpingo-oophorectomy (RRSO) significantly reduces the risk of ovarian, fallopian tube and breast cancer in BRCA mutation carriers and improves overall survival It is imperative to offer these patients who test positive for the BRCA1 mutation -2 the possibility of RRSO with possible hysterectomy. The sudden deletion of the ovarian follicular heritage brings with it a whole series of short-term disorders, mostly of the vasomotor type, or long-term cardiovascular and osteoporotic types. Therefore, in consideration of all these factors and to improve the quality of life of these women, the introduction of hormone replacement therapy (HRT) is necessary. BRCA mutation carriers undergoing RRSO can opt for a course of HRT after surgery, but there are concerns about enhancing their already increased risk of breast cancer. Notably, however, typical prescribed doses of estrogen (±progesterone) are considerably lower than physiological hormone levels in premenopausal women, and it would be difficult to significantly increase a BRCA carrier's risk of breast cancer since it is already extremely high. Armstrong et al. performed decision analysis on pooled epidemiological studies of women with BRCA1 and 2 mutations and concluded that HRT after RRSO was associated with a short-term benefit on life expectancy, up to age 50 years. In addition, some authors investigated whether the reduction in breast cancer risk conferred by RRSO in BRCA1 and 2 mutation carriers was impaired by post-RRSO HRT use. In this prospective cohort study, RRSO was significantly associated with reduced risk of breast cancer (hazard ratio [HR] = 0.40; 95% CI, 0.18 to 0.92), and use of HRT of any type after RRSO did not significantly alter the reduction in breast cancer risk associated with RRSO (HR = 0.37; 95% CI, 0.14 to 0.96). The authors concluded that short-term HRT use does not negate the protective effect of RRSO on subsequent breast cancer risk in BRCA 1 and 2 mutation carriers.
An important consideration in BRCA mutation carriers is that the majority of breast cancers that develop are estrogen/progesterone receptor negative (3), providing further reason to consider estrogen-based HRT. In a study of 456 BRCA mutation carriers by Eisen et al., 68% of BRCA-associated breast cancers were estrogen receptor negative, and there was no increased rate of cancer in those BRCA carriers on therapy hormone replacement. Interestingly, HRT use was associated with a significantly reduced risk of breast cancer (odds ratio 0.58, 95% CI 0.35-0.96) in this study.
The existing risk or diagnosis of breast cancer, as well as the decision to perform a concomitant hysterectomy at the time of RRSO, may influence the use of HRT in carriers of genetic mutations. A recent bulletin from the American College of Obstetrics and Gynecology states that reasons a physician might offer concurrent hysterectomy include
-the desire to completely remove the fallopian tubes due to a theoretical risk of fallopian tube cancer,
- to reduce the risk of pathology
endometrial in women who will be on tamoxifen therapy e
-simplification of HRT so that it can be taken
consider estrogen-only therapy (4).
Therefore, given the findings of the WHI study suggesting that estrogen therapy alone does not negatively impact breast cancer risk, a valid rationale for performing hysterectomy with RRSO would be to allow estrogen-only HRT if desired by the patient or the doctor.
REdiscovering BiOmarkers for the diagnosis and early tReatment respoNse in NEN. REBORN Study - NCT04464122.
The study aims to identify new markers for the diagnosis and follow-up of well-differentiated neuroendocrine neoplasms (NEN), in order to predict response to treatment before the classic morphological evaluation. All subjects enrolled in this observational, prospective, controlled study will undergo a screening visit (V0) and a baseline visit (V1). Patients affected by G1 and G2 well-differentiated NEN originating from the gastroenteropancreatic (GEP) tract will also undergo, after the start of medical therapy with somatostatin analogues or after surgery, a visit after 30 (V2) and 90 days (V3) and one year (V4) from V1.
The study includes a control group made up of patients referred to the benign endocrinopathies clinic of the Department of Experimental Medicine. The primary objective of the study is to evaluate the change in serum levels of the Tie2-soluble mediator after treatment with somatostatin analogues or surgery. The study also aims to investigate the following secondary objectives: quantification by flow cytometry of subpopulations of leukocytes and peripheral blood mononuclear cells (PBMC), evaluation of angiogenesis mediators and their receptors, evaluation of circular RNA extracted from tumor educated platelets (TEPs). Currently 16 patients with GEP-NEN G1 and G2 have been enrolled at the neuroendocrine neoplasms - NETTARE clinic and have carried out the investigations foreseen in V1, the results of which have been presented in a first work submitted for the published and currently under review. The first results concerning the characterization of circulating immune cells and their correlation with the levels of mediators of the Tie2/Angiopoietin pathway in patients with GEP-NEN G1 and G2 were presented by me as a poster at the European Congress of Endocrinology (ECE) 2022, held in Milan between 21 and 24 May 2022, and as an oral communication at the V National Congress of the SIE Oncological Endocrinology Club, held in Syracuse between 30 June and 2 July 2022.
The Mediterranean Ketogenic Diet as a possible sustainable intervention to boost the immune system of the metabolically unhealthy obese: achieving longer intervals between COVID-19 vaccines
Obesity and infection susceptibility are strictly and bidirectionally interconnected: patients with inflammatory disease or metabolic derangements such as obesity and T2D have been shown to be particularly susceptible to COVID-19, because of immune dysfunction, and the response to the vaccine is often suboptimal and rapidly waning over time, to the point that these fragile patients may need boosters very frequently to ensure adequate protection, with significant costs, contributing to the economic crisis caused by the pandemic. Obesity is characterized by an excess of adipose tissue. The adipose tissue is an endocrine organ and releases several adipokines which modulate inflammation (TNF, IL-6, MCP-1), insulin sensitivity and appetite (adiponectin, resistin, leptin), and Free Fatty Acids (FFAs). Excess circulating FFAs may represent one of the most important factors influencing insulin resistance (IR) and T2D in people with obesity. The adipokines TNFα, IL-6, and MCP-1 are involved in the induction of IR via upregulation of inflammation mediators.
The Mediterranean Diet is one of the most studied dietary patterns. The growing interest in the Mediterranean Diet arise from the evidence that highlighted the protective role of the Mediterranean model against different chronic diseases thanks to a quantitatively and qualitatively better consumption of food, rich in antioxidant and anti-inflammatory compounds. Foods comprised in the Mediterranean pattern contain high concentrations of polyphenol, carotenoids, vitamins, as vitamin C and E, folates and flavonoids: all these nutrients are well known to exert antioxidant and anti-inflammatory properties. Moreover, the type of fats consumed are unsaturated and polyunsaturated fatty acids (especially omega-6 and omega-3, contained abundantly in olive oil, fish and nuts as almonds, walnuts and pumpkins seeds): both omega-3 and omega-6-derived metabolites have important immune-regulatory functions. Besides these beneficial effects, the importance of the Mediterranean Diet is also conferred to a whole series of other socio-cultural, economic, and environmental benefits. In 2010, Mediterranean Diet has been recognized by UNESCO as an "intangible heritage of humanity" representing a set of skills, knowledge, practices, and traditions ranging from landscape to table; it has been also recognized as a sustainable diet. Indeed, the Mediterranean model has been shown to have a better ecological footprint compared with other dietary patterns and has therefore a smaller impact on the environment.
Nevertheless, this Mediterranean model, studied for the first time by Ancel Keys and associated with beneficial effects on the health and the environment, has nothing to do with the modern dietary pattern of Mediterranean countries and Italy specifically: indeed, the original Mediterranean Diet did not contemplate the current high levels of refined carbohydrates on which the typical Italian diet is based. With the view of inducing weight loss, a low-calorie dietary strategy based on high consumption of carbohydrates and low-fat content, in according to the modern Mediterranean dietary patter, yield only modest weight loss and suffer from low long-term compliance, compared to a Ketogenic approach which have been shown to be more effective, at least in the short to medium term.
For this reason, our study aims to connect the original Mediterranean Diet’s pivotal points with the effectiveness of the Ketogenic Diet on weight loss. By treating patients with metabolically unhealthy obesity with a Mediterranean style Ketogenic Diet, evaluating its impact on the immune response to the vaccine, we aim to identify a sustainable intervention, capable of preserving the ecosystem and biodiversity and at the same time allowing for longer intervals between COVID-19 booster doses. This in turn would have a significant impact on the pandemic related health costs, contributing to overcome the economic crisis.
Subjects are currently being enrolled among those accessing the Polyclinic Umberto I, Rome, undergoing a COVID-19 booster. The inclusion criteria are as follows: age ≥ 18 years old, stable body weight (less than 5 kg self-reported change during the preceding three months), intention to undergo vaccination, body mass index (BMI) ≥ 35 kg/m2 with at least one obesity-related complication or BMI ≥ 40 kg/m2 alone. The exclusion criteria are: immunodepression, use of medications known to impact the immune system, use of anti-obesity medications or others potentially affecting body weight or SGLT2 inhibitors, pregnancy, lactation, glomerular filtration rate (GFR) < 60 mL/min, insulin-dependent diabetes, or history of hyperuricemia. Data about demographic characteristics were collected through a structured interview. All the subjects are evaluated at baseline and after the vaccination. These subjects undergo a Mediterranean Ketogenic Diet and will be compared to those unwilling to undergo any dietary intervention (i.e. habitual diet).
The development of electrochemical biosensors for the determination of hormones and their metabolites.
The project is divided into two lines of research: i) the design and development of innovative sensors/biosensors for the monitoring of catecholamines; ii) the creation of new highly responsive, biocompatible and eco-sustainable electrode platforms. Some brain-related behaviors, such as stress, panic, anxiety, and depression are highly dependent on catecholamine levels. Therefore, being able to develop a reliable detection device capable of monitoring these levels in biological fluids, for early diagnosis, is of utmost importance. As part of this research, the first biosensor based on the enzyme tyrosinase (Tyr) and chitosan nanoparticles (ChitNPs) was developed to monitor the levels of Dopamine (DA), Epinephrine (EP) and Norepinephrine (NEP), in real samples of human urine. Optimization of the electrochemical platform was performed by enzymatic immobilization with 1-ethyl-3-(3-dimethylaminopropyl)-carbodimide (EDC) and N-hydroxysuccinimide (NHS), used as crosslinking agents. This biosensor made it possible to detect total catecholamine levels without interference, with a very low limit of detection (LOD) (0.17M for DA), excellent sensitivity, fast response time and good stability over time. The performance of the biosensor was subsequently evaluated in human urine samples, showing satisfactory results and, therefore, demonstrating the possible use of the proposed biosensor for the analysis of catecholamines in physiological samples.
The second study focused on the preparation of lignin/chitosan hybrid nanoparticles for the design of advanced, all-natural, biocompatible and environmentally sustainable biosensors. Lignin is the most abundant polyphenol in nature and is characterized by a complex chemical structure comprising phenylpropanoid units organized in a disordered sequence. The self-assembly of lignin into ordered nanoparticles (LNPs) is considered a promising alternative to traditional approaches, thanks to the improved physico-chemical, rheological and electrochemical properties as a consequence of the structural organization of the polymer's aromatic subunits. Lignin/chitosan hybrid nanoparticles (LNPs/CS) were thus synthesized using two procedures, including Layer-by-Layer (LbL) technology and Nanoprecipitation (Np). It has been demonstrated that the most effective LNPs/CS are produced by the Nanoprecipitation procedure, a practical and simple method for obtaining electrochemically active platforms both on a laboratory and pilot scale. The ease of preparation of the lignin/chitosan nanodevices, associated with a higher sustainability in terms of raw materials and a better electrochemical efficiency compared to commercial references suggests the possibility that the hybrid lignin/chitosan particles could represent an interesting alternative for the design of a new generation of electrode, selective and eco-sustainable platforms.
SECOND YEAR REPORTS
The main lines of research concerned:
New frontiers in the pathogenesis and management of comorbidities in the acromegalic patient
The goal of creating a device useful for screening and remote monitoring of the comorbidities of the acromegalic patient, through the evaluation of the main vital parameters and glycemic trend in patients suffering from acromegaly. With the aim of increasing the number of cases (final target 50 patients), in order to use the device also for the optimization of therapeutic schemes, and to minimize hospital admissions for these "fragile" patients, to start a multi-centre study. Potential implementation of the multiparametric device with the 24 h ECG holter. In view of the "clinical fragility" of these patients, who are affected by a 360 ° systemic pathology, we have decided to focus our attention, not only on the macroscopic aspects of the comorbidities of the acromegalic patient, but also on the microvascular and inflammatory aspects as a possible pathogenesis of organ damage, making use of the expertise of the Department of Translational and Precision Medicine (Clinical Immunology). All this with the aim of identifying, if possible, a common denominator, which can offer the basis for the creation of new pathogenetic and therapeutic frontiers in such complex patients and improve their quality of life. 20 patients with acromegaly were enrolled in this pilot study (7 males, 13 females), median age is 53 years (IQR 42–65 years), disease duration 12.5 years (IQR 6.5–21 years), and body mass index (BMI ) 24.8 kg/m2 (IQR 23.6-24.9 kg/m2) and 20 healthy controls matched for sex, age and BMI. The exclusion criteria used for both groups were: diagnosis of specific organ or systemic autoimmune disease, solid and/or haematological malignancies, metabolic diseases: overweight/obesity, diabetes, IGT/IFG, dyslipidemia, immunosuppressive therapy in the last 6 months and smoke. Two whole blood (WB) samples were collected to evaluate Interleukin IL 33 (IL33), serum Resolvin D1 (RvD1) levels and to identify the autoimmune profile (C3; C4; Antinuclear antibodies, ANA; SSa; SSb ; Sm; RNP; ScL70; Jo-1; Rheuma test, antibodies against citrullinated protein, anti-CCP), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR); blood count. Peripheral blood perfusion (PBP) of the dorsum of the hand was analyzed by laser speckle contrast analysis (LASCA) and Nailfold videocapillaroscopy (NVC) was performed for each patient.
Preliminary results suggest that IL 33 was significantly higher in patients with acromegaly than in healthy controls (p = 0.017) and RvD1 was significantly lower in patients with acromegaly than in healthy controls (p < 0.039). At LACA, mean peripheral perfusion (PBP) and Regions of Interest (ROI) I and ROI 3 were significantly higher in healthy controls than in acromegalic patients (p < 0.001; p 0.02; p 0.002, respectively). Abnormalities classifiable as nonspecific patterns were found on videocapillaroscopy in 20 patients (100%).
The study confirmed the presence of a subclinical inflammatory state associated with the increased IL33 concentration, which could contribute to the development of Cardiovascular Disease (CVD) in acromegaly. At the same time, acromegalic patients have a lower RvD1 concentration than healthy controls, which could be associated with a greater difficulty in resolving inflammatory processes. The relevant LASCA data on PBP and ROI I and 3 may correlate with lung perfusion in acromegalic patients; however this observation is still speculative and needs to be confirmed.
Future objectives: To expand the series to evaluate whether the different types of treatment can help improve the inflammatory state.
Novel biomarkers in type 2 diabetes and effect of exercise training: Bone Markers in Diabetes (BMD) study
The fracture risk in patients with type 2 diabetes mellitus (T2D) is higher than in the general population; however, in contrast to postmenopausal osteoporosis, it is associated with normal or increased bone mineral density (BMD) and low turnover.
The aim is to study the involvement of 5 new biomarkers (undercarboxylated osteocalcin (unOC), periostin, sphingosine 1-phosphate (S1P), VEGF (vascular endothelial growth factor) and GDF15 (Growth Differentiation Factor 15) in bone damage associated with T2D and, furthermore, to evaluate the impact of exercise.
The cross-sectional study design plans to measure these biomarkers in diabetic patients by comparing them with a group of osteoporotic patients (group C) and a healthy control group (non-diabetic non-osteoporotic subjects: group D). For our study, we will use patients already enrolled in the SWEET BONE study (“Study to Weigh the Effect of Exercise Training on BONE quality and strength in T2D”), who are required to take an additional sample for the dosage of the markers in question study. Patients with T2D are divided into group A, not physically active, and group B, physically active, which will have completed the 2 years of supervised and controlled physical exercise envisaged by the SWEET BONE protocol. Patients with osteoporosis should be active treatment naïve and the presence of secondary causes of osteoporosis should be excluded. All study participants will be male and female, aged between 65 and 75 years. The dosage of the biomarkers will be performed at the Bone Biochemistry Laboratory of the University of Sheffield (UK), directed by Prof Richard Eastell.
Study of GDF15, a marker of senescence, with regulatory activity on appetite and whose involvement in bone metabolism is still poorly understood. First of all, we conducted a first "ancillary" study which aimed to evaluate the impact of age and gender on GDF15 and its correlation with bone health parameters, in a population of healthy subjects divided into three age groups (Xtreme CT studies). The GDF15 is significantly higher in elderly subjects (over 70) and among men in this age group; however, we did not find significant correlations between GDF15 and BMD, BTMs, HRpQCT and hormones.
Subsequently, this marker was studied in a cohort of 1174 men and women from the Health ABC study, to evaluate its association with the risk of hip fracture: high levels of GDF15 are associated with a significant increase in the risk of hip fracture; however, this does not appear to be mediated by an association with BMD and action on bone, but by muscle component involvement and sarcopenia. As part of the Xtreme CT study, we also dosed VEGF, noting any association with the aforementioned bone health parameters.
Prospective longitudinal observational study ITACA (Impact of adrenal IncidenTalomas and possible Autonomous cortisol secretion on Cardiovascular and metabolic Alterations - NCT04127552).
The study aims to analyze the cardiovascular and metabolic alterations of patients with incidentally found adrenal masses and characterized by possible autonomous secretion of cortisol (subclinical hypercortisolism - PACS) compared with patients presenting with non-secreting adrenal masses (NFA) at baseline (T0), after 1 year (T1) and after 5 years (T2) of surveillance or adrenalectomy, following an internal algorithm based on the latest guidelines of the European Society of Endocrinology. The primary objective of the study is to evaluate with echocardiogram the change in indexed left ventricular mass in patients with pACS in T1 and T2 after surveillance or adrenalectomy. The study also aims to investigate any differences in baseline and in the following secondary objectives: cardiovascular parameters, anthropometric parameters, arterial stiffness, arterial pressure, alterations of hormonal and coagulation function, modifications of adrenal mass, quality of life, sleep, sexual function and the number and severity of infections. Currently 125 patients with adrenal incidentalomas have been enrolled in the adrenal pathology clinic (SMIP04) and have carried out the investigations foreseen at T0, the first results of which have been published in a first work (Sbardella E., Minnetti M., EJE 2018 ) which showed a higher prevalence of cardiac hypertrophy and diastolic dysfunction in patients with pACS.
The results concerning the infectious pathology (secondary outcome of the study) of patients with adrenal incidentalomas and pACS were recently published in the European Journal of Endocrinology (Minnetti et al. Susceptibility and characteristics of infections in patients with glucocorticoid excess or insufficiency: The ICARO tool; 2022 EJE). This study also contains the validation of the ICARO questionnaire, aimed at investigating the prevalence and severity of infections in patients suffering from endocrine pathologies, including Cushing's syndrome and adrenocortical insufficiency. Data on coagulation disorders in patients with adrenal incidentalomas were also presented by me in May at the Congress of the European Society of Endocrinology. As far as adrenal pathology is concerned, in the last year I have also worked as Sub-investigator of the phase III studies:
• CORT125134-455 (A Study of the Efficacy and Safety of Relacorilant in Patients With Endogenous Cushing Syndrome -GRACE)
• CORT125134-456 (Efficacy and Safety of Relacorilant in Patients With Cortisol-Secreting Adrenal Adenomas -GRADIENT)
I am also data manager of the multicenter study GONADIS promoted by the Italian Society of Andrology and Medicine of Sexuality SIAMS, aimed at describing gonadal, reproductive and psycho-sexological dysfunctions in male and female patients suffering from endocrine pathologies of adrenal and pituitary origin.
Validation of a new technology to assess body composition based on 3D photo-scan and bio-impedance
The project is focused on the assessment of body composition (CC) and aims to develop and test a new technological solution, the "Holistic Biometric Technology of Analysis of Human Body Composition" (BhOhB) which, through 3 digital photographs of the body, in an orthostatic position (front - side - back), evaluates not only the posture (already validated in the physiatric-rehabilitation area) but also the CC, which will be validated by interpolating and comparing its measurements with the body bioelectrical parameters (total body and segmental) measured by Bioelectrical impedance analysis (BIA) and whole body body composition analysis MOC-DEXA.
Materials and methods
Definition of technical measurement procedure on the basis of multidisciplinary comparison.
The images of several patients were collected as a test to identify the best markers to apply on the patients in order to verify the best method in terms of cost/benefit – time/quality measurement ratio. Through the multidisciplinary comparison with Prof. Cammarota of the Department of Mathematics and the BHOB technicians we have come to identify, for the algorithm definition phase, white bands to be positioned in a predefined way on the patient's body.
Enrollment and execution on all participants of:
MOC DEXA: gold standard instrumental examination for the definition of the composition of lean mass and fat mass at the level of tissues, cells, molecules and chemical elements through a low-intensity x-ray scan. The examination is carried out with a HOLOGIC machine in "WHOLE-BODY" scanning
Bioelectrical impedance analysis: a SECA 525 or Human-Im-Touch test will be carried out (the equipment is on loan for use by the Food Science Research Unit; declarations of conformity attached) with acquisition of electrical data [resistance (Rz, Ω), reactance (Xc, Ω), and phase angle (PA, °)] at frequencies 1.5, 50, 100 KHz according to ESPEN guidelines (Kyle, 2004). Calibration for all equipment is performed before each measurement session by comparing the values of Resistance (Rz) and Reactance (Xc) measured by the instrument by applying the electrodes to a circuit whose Rz and Xc values are known at 50 kHz ( phantom supplied by the manufacturer). The measurement is considered accurate if the deviation is within ± 10 Ohm for Rz and ± 5 Ohm for Xc;
Anthropometric measurements.
Circumference: Supra-elbow Root Arm Ankle Medial Calf Supra-patellar Root Thigh
Length: Arm Forearm Thigh Leg
BHOHB photography examination and positioning of anthropometric landmarks for digital measurements.
Retrospective study on clinical and ultrasound outcomes in a group of AFAB (Affirmed Female at Birth) transgender people receiving GATH gender-affirming hormone treatment
The ICD-11 (International Classification of Diseases 11th) defines gender incongruence as a mismatch between the gender that an individual identifies with and the sex assigned at birth. Gender incongruity is not itself considered a disease, it encompasses a broad spectrum of transgender people (binary and non-binary) within its definition. However, when the perceived discrepancy between sex at birth and perceived gender identity causes significant distress or disability, a diagnosis of gender dysphoria may be appropriate. (DSM5) These individuals' perceived distress manifests itself in a combination of anxiety, depression and irritability which may result in a strong desire to change their body through medical and/or surgical interventions. Standards of care for gender incongruity include the use of GAHT (Gender Affirmed Homonal Therapy) hormonal treatments ) to develop and maintain desired sexual characteristics. Hormone therapy for trans people AFAB (Assigned Female atBirth) is based on one of the available testosterone formulations, which include injections of parenteral esters (testosterone enanthate or undecanoate) or transdermal formulations. The hormone treatment aims to induce AFAB a modification of secondary sexual characteristics, such as increased production of hair on the body and face, lean body mass, interruption of the menstrual cycle, lowering of the tone of voice, redistribution of body fat, hypertrophy of clitoris and increased libido. The individual variability in treatment is rather heterogeneous and is strictly dependent on the type of molecule and dosage used, but also on an individual response which can be genetically and epigenetically determined.
Recent studies demonstrate how genetic polymorphisms can influence the sensitivity of the androgen receptor and the signal transduction mechanism, determining a greater or lesser response to hormonal therapy. The effect of androgen therapy on the female pelvic organs (uterus and ovaries) has not yet been evaluated in the literature; although in these patients there is a reduction, up to the complete disappearance of ovarian hormonal activity, the anthropometric variations of these organs are not known. Based on the lack of studies in the literature describing these changes in AFAB patients receiving androgen therapy, our study focused on the evaluation of the anthropometric change of the pelvic organs, through the use of transabdominal pelvic ultrasound, in a group of AFAB people with gender incongruence at time zero (T0), after 6 months (T6) and after 12 months ( T12) from the start of the GAT.
43 AFAB people with Gender Incongruence (transgender men) belonging to the Gender Dysphoria/Incongruence Clinic of the Department of Experimental Medicine (Umberto I Polyclinic - "La Sapienza" University) were recruited
All patients in the study took GAHT with testosterone enanthate 250 mg, injected im every 28 days. Pelvic ultrasound was performed at the Prenatal Diagnosis Center of the Department of Obstetrics, Gynecology and Urological Sciences of Sapienza University of Rome. The basic ultrasound evaluation was performed in the early follicular phase (3rd - 5th day of the menstrual cycle), according to ASRM-AIUM (ASRM, 2015; AIUM Practice Parameter for the Performance of an Ultrasound Examination of the Female Pelvis 2020). Ultrasound scans were all performed after adequate preparation with a transabdominal convex probe to avoid exacerbation of genital dysphoria. Uterine and ovarian volume were obtained using the volume formula of an ellipsoid: Volume (cm3) = length (DL) x depth (DAP) x width (DT) x 0.523. The volumes calculated for the uterus and ovaries for each patient were compared by examining the variations between T0, T6 and T12.
The mean age of the sample analyzed was 18.3 ± 0.5. Baseline ultrasound parameters were within the biological range for cisgender women. At T6 a significant decrease in uterus and ovarian volume, as well as endometrial thickness, was detected, which remained stable at T12.
Effects of progestin-only pill contAiNing DROspirenone 4 mg on MEtabolism anDRogens and sexual function: ANDROMEDa STUDY
This study was possible thanks to the establishment, from March 2021, at the Department of Experimental Medicine of Rome "La Sapienza", of a clinic dedicated to Gynecological Endocrinology, which constituted the "setting" of the research activity and of the recovery of data in this area, thanks also to the possibility of performing, in addition to the visit, an ultrasound examination of the pelvis and hormonal dosages with the use of mass spectrometry.
Drospirenone is a progestogen that entered the market in Italy in February 2020, in the formulation of 4 mg/day, as a progestogen-only contraceptive.
The literature data up to now indicate that the therapy of first choice in young women with hyperandrogenism/polycystic ovary syndrome is estrogen-progestogen (PE) therapy. However, in patients with contraindications to the use of estrogens (e.g. presence of known thrombophilia, migraine with aura, obesity, hypertension and habit of smoking in women >35 years, dyslipidemia, postpartum), progestogen-only therapy appears to be the only indicated choice3.
Until recently, the only progestogen-only pill was represented by Desogestrel 75 mg/day, which does not have a specific antiandrogenic activity and therefore, in most cases, does not bring benefits in patients with PCOS or clinical hyperandrogenism.
Drospirenone, on the other hand, is a molecule potentially endowed with anti-mineralcorticoid and anti-androgenic activity, but its efficacy in improving the clinical symptoms of hyperandrognism (hirsutism, acne, alopecia) and on metabolic parameters, in the absence of the estrogenic component (EE), has not yet been evaluated.
Study type: prospective longitudinal observational
Objectives: the primary objective of the study is to evaluate the anti-androgenic effects (clinical and biohumoral markers), metabolic effects and sexual function after 3, 6 and 12 months of using the Drospirenone-based contraceptive pill 4 mg/day.
The secondary objective is the evaluation of the serum levels of some miRNAs (mir155, mir27b, mir21, mir124a), which could be used as biomarkers of the response to therapy in patients with hyperandrogenism.
Materials and Methods: The study involves the enrollment of women of childbearing age (between 16 and 40 years of age) who are ineligible for combined estrogen-progestogen therapy, with signs of biohumoral and/or clinical hyperandrogenism.
Women whose clinical-anamnestic characteristics belong to categories 3 and 4 of the WHO classification "medical eligibility criteria for contraception" for the use of oral progestogen alone3 and women who have used other hormonal therapies in the last 3 months are excluded.
Each woman enrolled is evaluated at time 0' (baseline) and after 3, 6 and 12 months from the start of therapy with drospirenone 4 mg/day per os.
In particular, for each woman the following are evaluated at time 0' and at follow-up:
- anthropometric parameters (BMI, blood pressure, waist circumference, hip circumference)
- skin parameters of hyperandrogenism such as: acne/seborrhea (by The Global Acne Grading System (GAGS), androgenic alopecia (by Ludwing scale), hirsutism (by Ferriman-Gallwey scale)
- bihumoral parameters correlated with hyperandrogenism: testosterone, androstenedione, SHBG, DHEAS
- other hormonal parameters: FSH, LH, 17 beta estradiol, TSH, Ft3, Ft4, prolactin, ACTH, cortisol
- metabolic parameters: glucose, insulin, total and HDL cholesterol, triglycerides and transaminases
- assessment of sexual dysfunction, through the FSFI and SF-36 questionnaires
- menstrual cycle characteristics, ovarian characteristics on transvaginal pelvic ultrasound
- serum levels of mir155, mir27b, mir21, mir124a
Steroid hormones are analyzed both with immunometric methods and with mass spectrometry.
Preliminary results
The study is still ongoing: since March 2021, 18 patients have entered the study, of which 15 have completed the 3-month follow-up, 10 the 6-month follow-up and 4 the 12-month follow-up.
We therefore performed an analysis of the data at 3 and 6 months.
Preliminary results indicate that taking Drospirenone 4 mg/day did not change the anthropometric parameters (BMI, waist circumference, hip circumference) and blood pressure values at 3 and 6 months of follow-up.
As far as the hormonal profile is concerned, we observed a significant reduction in testosterone and androstenedione as early as 3 months, while the increase in SHBG was not statistically significant, demonstrating the fact that DRSP does not have a marked action on the stimulus to the synthesis of SHBG in the liver (unlike EE).
Furthermore, it is interesting to note how estradiol levels at 3 and 6 months remain around 40-50 pg/ml, typical values of the medium-proliferative phase, not lower than the values found when taking combined contraceptives with EE.
Finally, there was no significant variation in the thyroid, prolactin, adrenal axis, lipid, glucose and liver function settings.
The scales we used to evaluate the skin parameters of acne, hirsutism and alopecia showed a slight reduction in scores, although not yet statistically significant. This is in line with literature data regarding the anti-androgenic effects of combined contraceptives, which generally do not show results before 6 months of intake.
As regards sexual function, the analysis of the FSFI scores showed that this was not compromised, despite the significant reduction in total testosterone levels.
Furthermore, ultrasound pelvic evaluation showed that 60% of the patients had polycystic ovaries (> 20 follicles according to the new definition) at the baseline level (T0) and that this percentage remained unchanged at follow-up.
Finally, the evaluation of the menstrual pattern showed that 3 months after taking the DRSP about 50% of the patients presented frequent bleeding-spotting, while at 6 months the most frequent menstrual pattern was amenorrhea. In this context, counseling to the patient is of fundamental importance, defining how menstruation during hormonal contraception has no biological or clinical significance and that the slowing down of the frequency of menstrual flows induced by hormonal contraception has no negative effects on health, but that at the on the contrary, it can enhance the positive effects on women's health.
In conclusion, from the preliminary data analysis we can say that the intake of DRSP 4 mg/day:
- involves a significant reduction in testosterone and androstenedione already at 3 months
- has no significant effect on SHBG
- maintains estradiol levels at a medium proliferative level
- does not involve a significant modification of the metabolic, anthropometric, lipid and glucose parameters
- involves a slight reduction in the clinical parameters of hyperandrogenism (acne, hirsutism) even if not statically significant at 3 and 6 months
- does not lead to an alteration of sexual function
- results in an amenorrhea rate of 40% 6 months after the start of therapy.
Role of ovarian suppression during the treatment of patients with locally advanced cervical cancer: a prospective observational study
Cervical cancer is the fourth most frequent cancer in women, accounting for 6.6% of all female cancers.
In recent decades, more and more attention has been paid to the quality of life of cancer patients. In this specific case, cervical cancer is a neoplasm that occurs more in young women with a good life expectancy.
Thanks to the improvement in treatment and early diagnosis, an increase in the survival of patients with cervical cancer has been observed with greater attention to the quality of life of these patients.
In 40% of cases, cervical cancer occurs in patients before the age of 40, sometimes in women who have not completed their reproductive cycle.
In this context of patients, in addition to the reduced fertility linked to the medical and surgical treatment, we must underline the more dramatic and harmful effect of an iatrogenic menopause, with all the related symptoms (e.g. hot flashes, osteoporosis, cardiovascular diseases, 'humor).
Treatment options for locally advanced cervical cancer (LACC) (FIGO stages Ib2-IVa) include concurrent chemoradiation or neoadjuvant chemotherapy followed by radical hysterectomy and bilateral pelvic lymph node dissection, with or without ovarian removal. Both approaches can lead to premature ovarian failure.
Several studies have demonstrated the toxicity of chemotherapy and radiotherapy at the ovarian level as this is a tissue with a high proliferative potential.
The incidence of ovarian metastases in cervical cancer is different in the squamous histology where the incidence varies from 0.22% in the initial phase of the disease to 2.17% in LACC, on the contrary in non-squamous histology, in the case specific in adenocarcinoma, the risk of metastasis is 9.85% in LACC.
Therefore, in cervical cancers with squamous histology, ovarian conservation could be considered, after careful patient selection and accurate counselling, preserving carefully selected patients from the effects of iatrogenic menopause.
Primary objectives:
• evaluate the efficacy of GnRH-a associated with an oral estrogen-progestogen in the prevention of chemotherapy-induced POF
• identify hormonal changes through serum assays (FSH, estradiol and AMH)
• ultrasound evaluation (ovarian volume and antral follicle count) during the treatment process, before chemotherapy/radiotherapy (T0), after chemotherapy/radiotherapy (T1) and after surgery (T2).
Secondary objective: quality of life assessment of cancer patients.
THIRD YEAR REPORTS
Below are the main lines of research defined through the titles of the theses discussed:
• Precision nutrition in sarcopenic obesity: the COVID experience
• Natural history, endocrine complications and testicular dysfunction in Klinefelter syndrome and high-grade sex chromosome aneuploidies
• Characterization of patients with discrepant diagnoses for acromegaly new acquisitions on "micromegaly"
• Hormone replacement therapy in patients with previous gynecological malignancy
• Immune and metabolic profile in acromegalic patients and the impact of the current medical treatment. results from the promise study
• Effects of a ketogenic diet and an isocaloric balanced diet on quality of life, sleep and circadian rhythm: a randomized clinical trial.
• Epicardial adipose tissue and alterations of glucose metabolism as possible predictors of the severity of the sars-cov-2 infection
• Rediscovering biomarkers in for the diagnosis and early treatment response in NEN reborn study tutor