Projects and collaborations

Quantification of Venetoclax by HPLC-UV

The aim of the project was to develop a simple, rapid, and reliable method for the quantification of venetoclax. An analytical method was therefore developed and validated for the determination of the drug in human plasma using HPLC-UV with solid-phase extraction on cartridges. We are currently applying the developed method to various types of samples (plasma, red blood cells, and PBMCs) from patients with acute myeloid leukemia undergoing treatment with venetoclax, evaluating the method's suitability for clinical routine. The data observed so far suggest the usefulness of correlating venetoclax levels with clinical outcomes in order to gather information that could improve the management of treatment efficacy and toxicity. Data review for this project is still ongoing, as the number of patients currently receiving venetoclax is low. We are collecting additional data to build a statistically sound publication, taking into account the disaggregation of data by sex and gender.

Quantification of Catecholamines by HPLC-UV

The central role of catecholamines in neurodegenerative diseases is still poorly understood. These neurotransmitters play a fundamental role in assessing the functionality of the nervous system and related disorders; therefore, their quantification in biological fluids is of great importance. To this end, a useful method was developed and validated in accordance with FDA guidelines for the extraction and HPLC-UV analysis of catecholamines in various biological fluids. The method developed is simple, sensitive, and selective. So far, it has been applied to plasma and brain samples from mice. We are currently also working on developing a specific genetic analysis aimed at detecting potential polymorphisms involved in the etiology of neurodegenerative disorders. Monitoring catecholamine levels using HPLC-UV, coupled with genetic analysis of specific polymorphisms, could therefore help optimize treatment for neurodegenerative diseases. The work carried out so far in collaboration with NICO (Neuroscience Institute of the University of Turin) has enabled the sharing of results at national and international conferences and has led to the publication of two scientific articles in which age and sex variables were thoroughly investigated:

Thio-Metabolome: Potential Implications in Onco-Hematologic Diseases

Glutathione (GSH) is a tripeptide synthesized independently of ribosomal pathways. The γ-glutamyl cycle contains the metabolic pathway for its synthesis and degradation. Three of the six reactions in this cycle are linked to other metabolic pathways (EMP, TCA cycle, methylglyoxal), which are fundamental for understanding the systemic basis of its pathogenic mechanisms. Experimental evidence shows a strong correlation between GSH-dependent metabolism and the survival of tumor cells under apoptosis/oxidative stress. Additionally, according to the literature, graft-versus-host disease (GvHD) appears to have a direct correlation with plasma levels of cysteine/cystine, combined with a redox imbalance involving altered GSH/GSSG levels and upregulation of inflammatory cytokines. The potential therapeutic assessment of GSH could have significant clinical relevance for patients undergoing hematopoietic stem cell transplantation (HSCT) who develop GvHD. To this end, the group is working on an original project with multiple objectives. The primary objective is to identify prognostic/diagnostic biomarkers for the clinical monitoring of GvHD after HSCT. Evidence of metabolomic changes in this clinical context could have a significant impact on the clinical management of GvHD—an area that is currently underexplored. The secondary objective of the project is the internal validation of the analytical methods developed during the study, which are not currently available for routine clinical use. The goal is to make the analysis of these biomarkers available within a year as part of the Clinical Pharmacology Service’s test offerings. These would help clarify certain clinical aspects and improve the quality of care. The current project is being conducted on peripheral blood samples from HSCT patients using several approaches: immunomagnetic analyses, functional assays, metabolomic analysis via UPLC-MS/MS, genetic analysis on patient DNA, and in vitro culture experiments on patient-derived PBMCs. The project is currently in the data collection phase. The aim is to publish a scientific paper on the thio-metabolome by the end of 2024.

Determination of Glyphosate and Its Metabolite (AMPA)

Glyphosate, synthesized in 1974, was selected for commercial distribution as an herbicide due to its ability to selectively inhibit the shikimate pathway—an essential enzymatic route that supports plant growth. Considered safe in terms of environmental impact and toxicological effects on animals, with low production costs and high market demand, glyphosate quickly gained global use in extensive agriculture. It has been the most widely used herbicide over the past 40 years. Glyphosate is metabolized into aminomethylphosphonic acid (AMPA) by plants and in the soil through degradation processes. Since its initial introduction to the market, the effects of glyphosate on plants have been widely studied. However, except for cases of occupational exposure to high levels of glyphosate-based herbicides, early reports of human toxicity have been lacking. Scientific literature regarding the biological effects of glyphosate on humans remains controversial. In this context, establishing a simple and safe method for the extraction, identification, and quantification of glyphosate and AMPA in biological samples is of fundamental importance for investigating the potential pharmacodynamic and pharmacokinetic characteristics of these two molecules. Most of the methods described in the literature for determining these concentrations, however, are extremely complex, time-consuming, and do not provide acceptable quantification results. We have developed and validated a low-cost, simple, effective, and reproducible method for extracting and quantifying glyphosate and AMPA in various biological samples in order to define their potential biological effects. This work has led to the filing of a patent during the evaluation period, which was registered nationally and recently extended at the European level ("Procedure for determining glyphosate and aminomethylphosphonic acid in a sample." Chiara F et al., National application filed May 27, 2021; international extension May 26, 2022). This topic also opens the door to sex- and gender-specific considerations. There are, in fact, different effects on male and female fertility: glyphosate interacts with the estrogen receptor alpha, activating transcription of estrogen response elements, altering gene expression of the steroidogenic system and, consequently, pituitary homeostasis, which modifies the production of LH, FSH, progesterone, estradiol, and testosterone. Furthermore, glyphosate exposure differs significantly between men and women—traces of glyphosate have even been detected in feminine hygiene products such as tampons.

Transgender People and Healthcare

For the past year and a half, our research group has been focusing on data collection regarding transgender individuals who interact with healthcare systems, the challenges they face, and the possibilities for change. Recognizing the significant lack of crucial information needed to adequately represent trans* people within the healthcare system and to support the development of Gender-Specific Medicine, the questionnaire titled "Trans People's Health Census"* served as a starting point to begin addressing these gaps.