Cigarette smoke (CS) is responsible for cell injury, DNA damage and inflammation at airway due to an increased intracellular oxidative stress. STAT3/ERp57 pathway has been found upregulated by CS in airways and seems responsible for oncogenic proliferation of damaged cells. Conversely, upregulating Nrf2 protein and cannabinoid system has been shown to prevent airway and lung injury. The natural sesquiterpene ß-caryophyllene (CRY) and its metabolite ß-caryophyllene oxide (CRYO) have been highlighted to possess chemopreventive properties against CS-mediated airway damage, so suggesting their possible protective role for smokers. In the present project, CRY and CRYO will be studied for their protective properties against biochemical and epigenetic changes, induced by a sample of condensed cigarette smoke (CSC) in bronchial epithelial and lung cancer cells. Also, taking into account that smoking can increase the aggressiveness of pre-existing cancers, the ability of test substances to interfere with CSC-induced migration and metastatization of lung, breast and prostate cancer cells will be studied. The possible control of CSC-mediated airway inflammation by cannabinoid system will be studied too. At last, the inhibition of class I HDAC enzyme will be evaluated as possible epigenetic chemopreventive mechanism.
Cell migration and cytotoxicity will be measured by known methods, while DNA-damage will be evaluated by micronucleus assay. Inflammation will be monitored by measuring the cytokine levels, and known cannabinoid agonists and antagonists will enclosed to evaluate the possible control by the endocannabinoid system. The modulation of the ERp57/STAT3 and Nrf2 pathways, the levels of metastatic MMP2, BIRC5 and TPX2 proteins and the activity of I HDAC enzyme will be assessed by specific proteomic and genomic analysis.
The expected results allow to acquire new knowledge on smoke-related injuries and can suggest new targets for pharmacological therapies.
Present research project allows to obtain different results, which could be useful both for characterizing the chemopreventive properties of the sesquiterpenes under study and for identifying new alternative and pioneering strategies to counteract the damages due to active and/or second-hand smoke exposure. The major expected results are described below.
- Giving new scientific knowledge on the molecular and epigenetic mechanisms involved in smoke-mediated cell damage, which can be reflected on many cellular processes, including inflammation, DNA-damage, block of genome repair, precancerous lesions, oncogenic proliferation and metastatic progression of CSC-damaged cells.
- Studying the effects of current smoking on the survivor expectation of oncologic patients and the possible increase of the aggressiveness of pre-existing cancers, in order to give a guidance on the correct habits to be followed for favouring the chemotherapy effectiveness.
- Patenting and publication the obtained results in peer review and open access journals.
Taken together, these findings could increase our knowledge on the pathophysiology of smoke-related diseases and suggest new target for pharmacological therapies.
Smoking-related illnesses also represents a high cost for direct healthcare (long-term hospitalization, pharmacological therapies and loss of productivity) and induce worrying social consequences. In this context, searching for potential chemopreventive agents against tobacco smoke gives a strong social and economic impact to the present research project. Furthermore, although some aspects of smoke injury have been highlighted, the true molecular mechanisms are still unknown, so our results could give new scientific knowledge. The expected results may also enter into a process of technology transfer to pharmaceutical industry for the development and practical application of the research.
An outstanding aspect is the involvement of young researchers for the project, who will be trained and will acquire specialized skills for a future career.
The articulation of the research group allows to merge different skills (including pharmaco-toxicological, biochemical, and pathological) gained over the years by researchers, so increasing the scientific quality of the project and its possibility of success.
The PI, Dr. Di Sotto, possesses a consolidated expertise in the field of chemopreventive studies on natural substances, particularly terpenes, flavonoids and glucosinolates. In the previous years, she received some grants by Sapienza University (PI of University projects 2011 and 2013; co-investigator of the Un. project 2016 - PI, Mazzanti G.) for evaluating the possible chemopreventive role of some natural compounds against smoke, obtaining encouraging results. Recently, she also approached the study of the chemosensitizing properties of natural substances in combined treatment with known anticancer drugs, and also evaluate the effect of different treatment protocols on the chemotherapy effectiveness. This activity is, also, confirmed by her scientific publications in peer reviewed journals (32 papers; total citations 497; total Impact Factor 85.8; H-index 11) and by the abstracts of national and international conferences.
The PI can take advantages of specialized expertise for pharmaco-toxicological studies on natural substances, biochemical and histo-pathological investigations, due to consolidated intradepartmental scientific collaborations among Dept. of Physiology and Pharmacology, Dept. of Biochemical Sciences and Dept. of Anatomical, Histological, Forensic and Orthopedic Sciences.
The pharmaco-toxicological expertise of Prof. Mazzanti G. in addition to the pathological skills of Prof. Mammola will allow to better study the inflammation process induced by CS and the possible protective mechanisms of the sesquiterpenes tested. The involvement of the PhD student Dr. Carissimi from Dept. of Biochemical Sciences will support the study of different modulators of the inflammation induced by CS at airways and lung. The project will be also supported by Dr. Di Giacomo, a post-doc fellow researcher already employed at Dept. of Physiology and Pharmacology, with a consolidated expertise in studying the chemopreventive and chemosensitizing properties of natural substances.
This articulation represents an essential requirement for the project reliability and gives this team the characteristics of a forefront group for studying biologically active substances in the prevention and treatment of diseases.
Modern instruments (among which Spex FluoroMax spectrofluorimeter, MJ MiniOpticon Detection System, Leica DM5000B fluorescence microscope, Epoch microplate reader) and adequate laboratory infrastructures for cell culturing and treatment are available at departments involved in this research, so increasing the reliability of the project.