Periodic Reporting for period 2 - INTEGRATA (Integrating chemical and biological approaches to target NAD production and signaling in cancer) Reporting period: 2020-11-01 to 2023-04-30 Summary of the context and overall objectives of the project INTEGRATA aimed at providing a group of early stage researchers (ESRs) with a highly interdisciplinary and intersectoral training in drug discovery, focusing specifically on NAD production and on NAD/nucleotide signaling as targets for the development of new cancer therapeutics1. This given that interrupting NAD production, or specifically targeting enzymes from the NAD/nucleotide signaling apparatus, is considered a very appealing and novel approach in oncology and one of the most promising areas of investigation for the development of new drugs2. Biological targets in focus. INTEGRATA aimed at exploiting the therapeutic potential of nicotinamide phosphoribosyltransferase (NAMPT) as a target to inhibit, but also of additional enzymes from the NAD-biosynthetic and NAD/nucleotide signaling apparatus, for which mounting evidence indicates a role in cancer. These included nicotinic acid phosphoribosyltransferase (NAPRT), SIRT6, CD73, transient receptor potential cation channel, subfamily M, member 2 (TRPM2), and the ADP sugar-pyrophosphatase NUDT5.On these grounds, INTEGRATA pursued the following scientific and technological objectives:• To develop new agents for interfering with NAD biosynthesis in cancer cells [NAMPT, NAPRT, inhibitors and Ab drug conjugates (ADCs) to target NAMPT inhibitors to multiple myeloma (MM), B-cell lymphoma and AML cells], neutralizing monoclonal antibodies (mAb) against eNAMPT and eNAPRT and inhibitors of SIRT6, NUDT5, CD73, and TRPM2.• To perform an extensive testing of the newly generated NAD biosynthesis and NAD/nucleotide-signaling inhibitors in cultured cells and in in vivo cancer models, including models for assessing angiogenesis and cancer cells’ metastatic spread. • To conduct pre-formulation/fingerprinting, formulation and pharmacokinetics (PK) studies with the newly produced NAD biosynthesis and NAD/nucleotide-signaling inhibitors.• To define anticancer activity and potential toxicities of the newly generated agents, as well as of a dietary approach based on a NA-free diet w/ or w/o antibiotics (to reduce NAPRT activity) in relevant in vivo cancer models. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far The work performed in the last 24 months and the results achieved so far are detailed below:• The ESRs from the INTEGRATA consortium successfully produced new inhibitors with the ability to reduce intracellular NAD+ stores, such as NAMPT and NAPRT inhibitors, as well as Ab drug conjugates (ADCs) to target NAMPT inhibitors to multiple myeloma cells. Neutralizing monoclonal antibodies (mAb) against eNAMPT as well as NAD/nucleotide signaling inhibitors, such as NUDT22 and TRPM2 inhibitors have also been developed and tested. • The newly generated NAD biosynthesis and NAD/nucleotide-signaling inhibitors were extensively tested in cultured cells and in in vivo cancer models. Pre-formulation/fingerprinting, formulation and pharmacokinetics studies were performed with SIRT6 inhibitors and with selected NAMPT inhibitors.• Finally, anticancer activity and potential toxicities of a dietary approach based on a NA-free diet w/ or w/o antibiotics (to reduce NAPRT activity) in relevant in vivo cancer models have also been determined.Overall, the project achieved its goals and had a major scientific impact as shown by the numerous scientific articles that have been published so far or that are in preparation or in press. The INTEGRATA ESRs performed numerous dissemination activities [such as taking part in international scientific meetings or in public events such as the “Notte dei Ricercatori” (“Researchers’ night”)] and attended courses on topics such as “Scientific communication and presentation of scientific data”, “Business and financial analysis in drug discovery, patenting, entrepreneurship, regulatory affairs”, etc. The latter activities helped building their scientific background and increasing their opportunities for a future, successful career in science. Therefore, the project has achieved significant scientific advances and has successfully trained fourteen young scientist in the field of NAD+ biology and cancer metabolism through a truly interdisciplinary approach. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) The originality and innovation of INTEGRATA’s research programme primarily lied in its ambition to translate strong skills in biochemistry and knowledge of NAD production and signaling into applied cancer research and cancer drug discovery, but also in providing its PhD students with knowledge and skills required in the technology transfer process and that are typical of successful biomedical enterprises. This vision was implemented in leading European labs, SMEs and industries in an overarching training programme, which prepared our ESRs to take the lead in the innovation process in cancer drug discovery ongoing in Europe and worldwide. The technological objectives of INTEGRATA have been innovative inasmuch as they aimed at delivering inhibitors for highly promising targets, for which no clinically applicable drug has been identified yet (e.g. NAPRT, SIRT6, TRPM2 and NUDT22) and that could thus become active cancer drugs in the near future. A strong point of the project was also the involvement of four clinical centers, two SMEs and of an industry in it. Oncologists and haematologists were actively involved in the training of the INTEGRATA ESRs, sharing their experience in the clinical hurdles of the different types of cancer and updating the ESRs on the state-of-the-art of cancer drug treatment. In addition, their involvement was also useful to facilitate the transition of newly identified active agents into future clinical studies. The non-academic partners provided their expertise and infrastructures in addition to training the ESRs within successful entrepreneurial environments. The partners worked in strict collaboration, with frequent secondments of the fellows to other partners, thus creating an optimal synergy for reaching the common research training goals. The plan of fellows’ secondments were the basis for ensuring the effectiveness of networking and they paved the way for a successful future career of the fellows with opportunities in both academic and non-academic sectors. Importantly, the ultimate goal of our training programme, i.e. the creation of a core group of experts with interdisciplinary skills in NAD biology, cancer metabolism and drug development, went well beyond current programmes and led to a broader approach to cancer treatment that is expected to attain a deeper vision of the problem, and to propose valid, new therapeutic solutions. Project logo
