The main objective of the project has been successfully accomplished as the proteasome machinery targets were identified using global proteomics and transcriptomics in a panel of 8 cell lines (6 cancer cell lines and 2 multiple myeloma cell lines) – and a map was made of molecular pathways regulated by the proteasome in the studied cells (see Figure 1 attached). The comparison of myeloma and cancer global proteome changes upon proteasome inhibition allowed for identification of 44 proteins/genes across 20 pathways which were candidate proteasome targets providing resistance to Carfilzomib in cancers vs. multiple myeloma. The following validation and siRNA screen allowed to focus on molecular chaperones, autophagy, endocytosis and unfolded protein response pathways - with molecular chaperones and their co-chaperones having a pivotal role in all the listed pathways. In consequence, tested specific molecular chaperone inhibitors and selected co-chaperone inhibitors provided the additive effect in killing cancer cells with Carfilzomib, allowing to bypass resistance to this inhibitor. Thus, the project, thanks to the global identification of the proteasome machinery targets, has allowed finding drug targets and preclinical protocols for treatment of colon, lung and pancreatic cancer cells, which rely on increasing the efficiency of the proteasome inhibitor Carfilzomib.
All the major points of the expected exploitation and dissemination have been successfully accomplished:
1. The project greatly helped the project’s Researcher to establish (formally in October 2018) a laboratory consisting currently of a staff of 5-10 postdocs, PhD and MSc students - Laboratory of Human Disease Multi-omics at MMRC PAS, Warsaw, Poland. The laboratory is successfully using the workflow established in this project for other studies.
2. The progress and methods were communicated during seminars in the Beneficiary institution and collaborating institutions – e.g. University of Warsaw (
https://cent.uw.edu.pl/en/blog/dawid-walerych-mossakowski-medical-research-centre-warsaw/(se abrirá en una nueva ventana)) or International Institute of Molecular and Cell Biology in Warsaw (
https://www.iimcb.gov.pl/en/press-office/news/seminars?start=50(se abrirá en una nueva ventana)).
Especially successful was the open lecture/workshop series “Cancer Fight Research Club” (
https://twitter.com/FightCancerClub(se abrirá en una nueva ventana)https://www.facebook.com/CancerFightResearchClub/(se abrirá en una nueva ventana)) organised at the Beneficiary institute (September 2019 – February 2020). This allowed to invite known cancer researchers from Poland and EU countries to give lectures, talk to them about the project and further collaborate in it. These meetings attracted audience from many science and education institutions, also non-professional audience – serving science popularization.
3. Two conference announcements related to the project were done: A poster during the 8th Mutant p53 Workshop in Lyon, France, 13-18 May 2019 (https://p53.iarc.fr/download/8thp53MutantWorkshopBook.pdf - the proteasome gene transcription and activity was a readout of the mutant p53 oncogenic activity) and a poster and a short talk were done at 13th Central and Eastern European Proteomic Conference, 23-25.09.2019 Ustoń, Poland (
http://cd.io.gliwice.pl/ceepc13/(se abrirá en una nueva ventana)).
4. One review manuscript was published concerning in part the proteasome machinery in cancer (
https://www.mdpi.com/2072-6694/12/6/1532(se abrirá en una nueva ventana)). The main project’s research manuscript will be submitted in the next months when the mechanism studies in compensation of the proteasome inhibition are complete, and the therapeutic tests in vivo will be completed (both are ongoing and were not listed as project tasks – they will allow to support the results better and publish them in a higher impact journal).
5. The experience on mass spectrometry and proteomics/transcriptomics data analysis gained during the project, especially during the Secondment, was indispensable for the project, extremely useful other projects and generally – for the
