0585842 - BTÚ 2025 RIV GB eng J - Článek v odborném periodiku
Mierzwicka, Joanna Maria - Petroková, Hana - Kafkova, L. R. - Kosztyu, P. - Černý, Jiří - Kuchař, Milan - Petrík, M. - Bendová, K. - Krasulova, K. - Groza, Yaroslava - Vaňková, Lucie - Bharadwaj, Shiv - Panova, Natalya - Křupka, M. - Škarda, J. - Raška, M. - Malý, Petr
Engineering PD-1-targeted small protein variants for in vitro diagnostics and in vivo PET imaging.
Journal of Translational Medicine. Roč. 22, č. 1 (2024), č. článku 426. ISSN 1479-5876. E-ISSN 1479-5876
Grant CEP: GA MŠMT(CZ) EF18_046/0015974
Výzkumná infrastruktura: CIISB III - 90242
Institucionální podpora: RVO:86652036
Klíčová slova: virus-neutralizing sera * cd8 t-cells * pd-1 * antibody
Obor OECD: Pathology
Impakt faktor: 6.1, rok: 2023 ; AIS: 1.446, rok: 2023
Způsob publikování: Open access
Web výsledku:
https://translational-medicine.biomedcentral.com/articles/10.1186/s12967-024-05210-xDOI: https://doi.org/10.1186/s12967-024-05210-x

Background Programmed cell death 1 (PD-1) belongs to immune checkpoint proteins ensuring negative regulation of the immune response. In non-small cell lung cancer (NSCLC), the sensitivity to treatment with anti-PD-1 therapeutics, and its efficacy, mostly correlated with the increase of tumor infiltrating PD-1+ lymphocytes. Due to solid tumor heterogeneity of PD-1+ populations, novel low molecular weight anti-PD-1 high-affinity diagnostic probes can increase the reliability of expression profiling of PD-1+ tumor infiltrating lymphocytes (TILs) in tumor tissue biopsies and in vivo mapping efficiency using immune-PET imaging.Methods We designed a 13 kDa beta-sheet Myomedin scaffold combinatorial library by randomization of 12 mutable residues, and in combination with ribosome display, we identified anti-PD-1 Myomedin variants (MBA ligands) that specifically bound to human and murine PD-1-transfected HEK293T cells and human SUP-T1 cells spontaneously overexpressing cell surface PD-1.Results Binding affinity to cell-surface expressed human and murine PD-1 on transfected HEK293T cells was measured by fluorescence with LigandTracer and resulted in the selection of most promising variants MBA066 (hPD-1 KD = 6.9 nM, mPD-1 KD = 40.5 nM), MBA197 (hPD-1 KD = 29.7 nM, mPD-1 KD = 21.4 nM) and MBA414 (hPD-1 KD = 8.6 nM, mPD-1 KD = 2.4 nM). The potential of MBA proteins for imaging of PD-1+ populations in vivo was demonstrated using deferoxamine-conjugated MBA labeled with 68Galium isotope. Radiochemical purity of 68Ga-MBA proteins reached values 94.7-99.3% and in vitro stability in human serum after 120 min was in the range 94.6-98.2%. The distribution of 68Ga-MBA proteins in mice was monitored using whole-body positron emission tomography combined with computerized tomography (PET/CT) imaging up to 90 min post-injection and post mortem examined in 12 mouse organs. The specificity of MBA proteins was proven by co-staining frozen sections of human tonsils and NSCLC tissue biopsies with anti-PD-1 antibody, and demonstrated their potential for mapping PD-1+ populations in solid tumors.Conclusions Using directed evolution, we developed a unique set of small binding proteins that can improve PD-1 diagnostics in vitro as well as in vivo using PET/CT imaging.
Trvalý link: https://hdl.handle.net/11104/0355227