The main areas of interest of our laboratory are:
- study of amyloidogenic proteins and identification of new therapeutic strategies for diseases characterized by amyloid accumulation (i.e. Alzheimer's disease, prion diseases, peripheral amyloidosis);
- development and application of innovative analytical assays to measure blood concentrations of therapeutic antibodies and anti-drug antibodies. Therapeutic drug and immunogenicity monitoring (TDIM) is important in guiding therapy with biologics, to permit objective decisions for the management of non-responders and reduce unnecessary interventions with these expensive treatments.
- development of new analytical assays for drugs, endogenous biomarkers, and biomolecules in general. These studies involve pharmacodynamic aspects, particularly the interactions between molecules through Surface Plasmon Resonance, as well as pharmacokinetic aspects using different analytical methods, including mass spectrometry. The laboratory is part of the “Centro di Analisi Biomedica e Genomica Clinica” (CABGeC) of the Istituto Mario Negri and, as such, it follows a certified Quality Management System UNI EN ISO 9001 with the following scope “Provision of biomedical analysis for clinical a diagnostic research in the clinical field”, in compliance with the current regulation applicable to pharmacokinetic studies in humans (Determina AIFA n. 809/2015).
Alzheimer’s disease and other amyloidoses
Alzheimer's disease, prion diseases and some peripheral amyloidosis are a group of diseases characterized by the formation of pathological protein aggregates (amyloids). Our group investigates the properties of these amyloidogenic proteins to identify new potential targets and therapies. In particular, we have developed innovative methods for the selective recognition, and analysis, of the most toxic protein aggregates, termed oligomers. We are applying these approaches to: i) characterize the molecular mechanisms underlying oligomer toxicity; ii) develop new tests for an early diagnosis, to identify patients who could most benefit from these therapies; iii) identify compounds capable of preventing oligomer toxicity. As regards this latter point, we are starting new projects to evaluate the properties of extracts or active principles obtained from medicinal plants. Moreover, we are exploring a new potential mechanism of action of doxycycline, a well-known antibiotic that counteracts the cognitive decline in mice models of Alzheimer’s disease through an anti-inflammatory effect. In particular, after a detailed characterization of the pharmacokinetic profile and brain distribution of doxycycline, we are currently investigating the inhibitory effect of the drug on PARP-1, an enzyme involved in the neuroinflammatory processes induced by amyloid aggregates.
Search for new drugs to fight SARS-CoV-2 infection
Our laboratory is participating in a project aiming to design new molecules potentially useful to prevent SARS-CoV-2 infection, responsible for the COVID-19 epidemic. In particular, the study aims to identify molecules capable of blocking the entry of the coronavirus into cells, which occurs through the interaction between the viral "Spike" protein and the ACE-2 receptor present on the surface of human cells. Our laboratory is conducting in vitro studies that allow to measure the binding capability of the new molecules to the ACE-2 protein, which hinders the complex formation between the Spike and ACE-2 protein.
New tests for personalized therapies with monoclonal antibodies
Many monoclonal antibodies are being approved in the clinical practice for their therapeutic benefit. Unfortunately, some patients do not respond, probably because of the onset of anti-drug antibodies that counteract the therapeutic activity. In our laboratory, we have developed a new test that uses Surface Plasmon Resonance technology, to simultaneously measure the drug and anti-drug antibodies in the blood of patients. This is usually not possible using other techniques (e.g. ELISA assays). We are currently analyzing blood samples of a large number of patients in order to unveil different properties of anti-drug antibodies and to determine their relevance for the response to therapy. The knowledge of these data for each patient will allow doctors to customize and optimize the therapy. With this in mind, we also plan to develop, in collaboration with engineering departments, new miniaturized and portable devices to allow rapid, easy and cheap point-of-care analysis.
Identification of molecular mechanisms underlying the brain damage induced by cardiac arrest
Brain injury represents a leading cause of mortality and long-term neurological disability in patients successfully resuscitated from cardiac arrest. Data obtained in the Department of Cardiovascular Medicine, in collaboration with our lab, revealed that a metabolic pathway is activated immediately after cardiac arrest, called kynurenine pathway (KP), which is associated with severity of post–cardiac arrest shock, early death, and poor long-term outcome. We additionally obtained preliminary data in a rodent model of cardiac arrest showing alteration of KP metabolites in the brain. Since some of these metabolites have important effects on neurons, we are currently evaluating their role in the cascade of molecular events underlying the cardiac arrest-induced brain injury. The KP could thus represent a novel pharmacological target useful to reduce morbidity and mortality in patients after cardiac arrest.
New tests for the identification of molecules with anti-ischemic activity
Ischemic stroke is the leading cause of death and permanent disability. Despite recent substantial progress in prevention and management, its successful treatment remains a large unmet medical need. Studies carried out in the Laboratory of Inflammation and Nervous System Diseases, in collaboration with our lab, suggested the pivotal role of mannose-binding lectin (MBL), one of the recognition molecules of the lectin complement pathway, in brain ischemic injury. These data support the hypothesis that MBL inhibition may be a relevant therapeutic target in humans. Our group has developed new methods based on Surface Plasmon Resonance to identify molecules able to counteract the detrimental effects of lectins. These compounds have neuroprotective activity in animal models undergoing ischemia, suggesting a promising treatment strategy of ischemic brain stroke. We are currently involved in a European project whose general aim is the development of glyco-nanoparticles suitable for applications in advanced nanomedicine. Within this project, our work will focus on the identification of nanoparticales able to interact with – and inhibit – the specific lectins responsible of ischemic damage.
Analysis of potential biomarkers in patients with septic shock
Septic shock is a systemic inflammatory response to infection characterized by profound circulatory abnormalities, leading to death in about 50% of patients.. Thus, it is crucial to identify and characterize biomarkers representative of the syndrome progression in order to optimize the therapy improving survival. In collaboration with the Department of Cardiovascular Medicine, we have developed and applied new analytical methods to measure the blood concentrations of three biomolecules in patients participating in the ALBIOS clinical trial. The three biomolecules, Syndecan-1, Sphingosine-1-phosphate and VE-cadherin, are important for maintaining the integrity and permeability of blood vessels, and can play a key role in the pathophysiological processes that follow septic shock. The study confirmed that the concentrations of the three biomolecules are related to the severity of the clinical situation and short-term survival.
Multiple Sclerosis: blood concentrations of new drugs
Multiple Sclerosis (MS), is the most common cause of neurological disability. The recent approval of new oral therapies offer exciting new opportunities to optimize treatment outcomes. We have developed and validated two new analytical methods following the appropriate international guidelines for the measurement of blood concentrations of two oral drugs approved for MS. In collaboration with the "Carlo Besta" Neurological Institute of Milan we monitor their blood levels, which will provide useful indications to interpret the clinical response of patients and for a better use of these drugs.
International Consensus on Cardiopulmonary Resuscitation.