Počet záznamů: 1
Engineering PD-1-targeted small protein variants for in vitro diagnostics and in vivo PET imaging
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SYSNO ASEP 0585842 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Engineering PD-1-targeted small protein variants for in vitro diagnostics and in vivo PET imaging Tvůrce(i) Mierzwicka, Joanna Maria (BTO-N)
Petroková, Hana (BTO-N) RID, ORCID
Kafkova, L. R. (CZ)
Kosztyu, P. (CZ)
Černý, Jiří (BTO-N) RID, ORCID
Kuchař, Milan (BTO-N) RID, ORCID
Petrík, M. (CZ)
Bendová, K. (CZ)
Krasulova, K. (CZ)
Groza, Yaroslava (BTO-N) ORCID
Vaňková, Lucie (BTO-N) ORCID
Bharadwaj, Shiv (BTO-N)
Panova, Natalya (BTO-N)
Křupka, M. (CZ)
Škarda, J. (CZ)
Raška, M. (CZ)
Malý, Petr (BTO-N) RID, ORCIDCelkový počet autorů 17 Číslo článku 426 Zdroj.dok. Journal of Translational Medicine - ISSN 1479-5876
Roč. 22, č. 1 (2024)Poč.str. 19 s. Jazyk dok. eng - angličtina Země vyd. GB - Velká Británie Klíč. slova virus-neutralizing sera ; cd8 t-cells ; pd-1 ; antibody Vědní obor RIV FP - Ostatní lékařské obory Obor OECD Pathology CEP EF18_046/0015974 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Výzkumná infrastruktura CIISB III - 90242 - Masarykova univerzita / Středoevropský technologický institut Způsob publikování Open access Institucionální podpora BTO-N - RVO:86652036 UT WOS 001214836800003 EID SCOPUS 85192176364 DOI https://doi.org/10.1186/s12967-024-05210-x Anotace 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. Pracoviště Biotechnologický ústav Kontakt Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Rok sběru 2025 Elektronická adresa https://translational-medicine.biomedcentral.com/articles/10.1186/s12967-024-05210-x
Počet záznamů: 1