In Vitro Testing of the Virus-Like Drug Conjugate Belzupacap Sarotalocan (AU-011) on Uveal Melanoma Suggests BAP1-Related Immunostimulatory Capacity.

Autor:	Ma S; Department of Ophthalmology, Leiden University Medical Center (LUMC), Leiden, The Netherlands., Huis In't Veld RV; Department of Radiology, Leiden University Medical Center (LUMC), Leiden, The Netherlands.; Department of Immunology, Leiden University Medical Center (LUMC), Leiden, The Netherlands., Houy A; Inserm U380, DNA Repair and Uveal Melanonoma (D.R.U.M.), Equipe labelliseé par la Ligue Nationale Contre le Cancer, Institut Curie, PSL Research University, Paris, France., Stern MH; Inserm U380, DNA Repair and Uveal Melanonoma (D.R.U.M.), Equipe labelliseé par la Ligue Nationale Contre le Cancer, Institut Curie, PSL Research University, Paris, France., Rich C; Aura Biosciences, Inc., Cambridge, Massachusetts, United States., Ossendorp FA; Department of Immunology, Leiden University Medical Center (LUMC), Leiden, The Netherlands., Jager MJ; Department of Ophthalmology, Leiden University Medical Center (LUMC), Leiden, The Netherlands.
Jazyk:	angličtina
Zdroj:	Investigative ophthalmology & visual science [Invest Ophthalmol Vis Sci] 2023 Jun 01; Vol. 64 (7), pp. 10.
DOI:	10.1167/iovs.64.7.10
Abstrakt:	Purpose: The virus-like drug conjugate belzupacap sarotalocan (AU-011), currently under clinical investigation for first-line treatment of primary uveal melanoma (UM), shows enhanced tumor specificity by targeting heparan sulfate proteoglycans (HSPG). Such a treatment may potentially lead to systemic immune responses. We studied the potential of AU-011 treatment to induce immunogenic cell death as the first step to induce systemic immunity. Methods: We determined binding and uptake of AU-011 in ten primary and metastatic UM cell lines. The subcellular location of AU-011 was assessed by fluorescence microscopy. Following light activation (wavelength 690 nm) of AU-011, the half-maximal effective concentration (EC50) of AU-011 treatment and exposure of damage-associated molecular patterns (DAMPs) were assessed using flow cytometry. DAMPs were measured by RNAseq. Results: Fluorescence microscopy revealed most of the AU-011 was present in the cytoplasm. AU-011 binding and uptake by UM cells increased over time, with a lower uptake in BAP1-negative than in BAP1-positive cell lines. AU-011 activation induced cell death across all UM cell lines with EC50 values at picomolar concentrations. The AU-011 concentration and total light dose (J/cm2) were the most important parameters for the observed cytotoxicity. Finally, light-activated AU-011 induced exposure of DAMPs calreticulin (CRT) and HSP90. CRT exposure by light-activated AU-011 as well as CRT RNA exposure were lower in BAP1-negative compared to BAP1-positive UM cell lines. Conclusions: AU-011 treatment at low picomolar range induces immunogenic cell death in all 10 UM cell lines. The in vitro cytotoxicity was accompanied by exposure of DAMPs (HSP90 and CRT), suggesting AU-011 may contribute to the development of systemic immunity and be a suitable candidate for combination with immunotherapy in vivo. AU-011 treatment was more effective against BAP1-positive cell lines, with a lower EC50 and higher CRT exposure.
Databáze:	MEDLINE
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