Engineering PD-1-targeted small protein variants for in vitro diagnostics and in vivo PET imaging

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F24%3A00585842" target="_blank" >RIV/86652036:_____/24:00585842 - isvavai.cz</a>

Alternative codes found

RIV/61989592:15110/24:73627271 RIV/61989592:15640/24:73627271

Result on the web

<a href="https://translational-medicine.biomedcentral.com/articles/10.1186/s12967-024-05210-x" target="_blank" >https://translational-medicine.biomedcentral.com/articles/10.1186/s12967-024-05210-x</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1186/s12967-024-05210-x" target="_blank" >10.1186/s12967-024-05210-x</a>

Result language

angličtina

Original language name

Engineering PD-1-targeted small protein variants for in vitro diagnostics and in vivo PET imaging

Original language description

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.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

30109 - Pathology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2024

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Translational Medicine

ISSN

1479-5876

e-ISSN

1479-5876

Volume of the periodical

22

Issue of the periodical within the volume

1

Country of publishing house

GB - UNITED KINGDOM

Number of pages

19

Pages from-to

426

UT code for WoS article

001214836800003

EID of the result in the Scopus database

2-s2.0-85192176364