A human receptor occupancy assay to measure <scp>anti‐PD</scp>‐1 binding in patients with prior <scp>anti‐PD</scp>‐1

Junker, Fabian; Gulati, Pratiksha; Wessels, Uwe; Seeber, Stefan; Stubenrauch, Kay-Gunnar; Deak, Laura Codarri; Markert, Christoph; Klein, Christian; Teixeira, Priscila Camillo; Kao, Henry

doi:10.1002/cyto.a.24334

Cited by 12 publications

(8 citation statements)

References 52 publications

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“…As described previously [41], healthy donor blood was collected in sodium heparin (NaHep) tubes (BD), diluted with PBS (Gibco, 1:1) and transferred to Leucosep tubes (Greiner Bio‐One) pre‐filled with Ficoll‐Paque Plus (GE Healthcare). Tubes were centrifuged without brake (1000 g, 10 min, RT).…”

Section: Methodsmentioning

confidence: 99%

Barcoding of live peripheral blood mononuclear cells to assess immune cell phenotypes using full spectrum flow cytometry

Junker

Teixeira

2022

Cytometry Pt A

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Barcoded flow cytometry is a multiplexing technique allowing for the simultaneous acquisition of cells from different donors or experimental conditions in a highthroughput manner. This approach allows to synchronize acquisition of samples and reduce variance introduced through the operator or technical platform. However, to date, only very few flow cytometry barcoding protocols have been developed, which often suffer from technical limitations. Here, we developed a novel barcoding protocol for a full-spectrum flow cytometry platform. We developed a 21-color immunophenotyping assay for up to 20 different samples analyzed simultaneously with comparable variance between repeated single-tube acquisition and postde-multiplexing. Barcoding offers great potential in parallelizing the analysis of complex cell populations such as peripheral blood mononuclear cells (PBMCs). Consequently, we assessed the performance of our method in situations where PBMCs were challenged with phytohaemagglutinin (PHA), a strong mitogen and broad activator of B cells and T cells, and superantigen Staphylococcus enterotoxin B (SEB) that has been reported to induce polyclonal T cell activation. PBMCs were either barcoded before pooled challenge or challenged individually pre-barcoding. Our final workflow included pooled immunophenotyping followed by machine learning aided single-cell data analysis and enabled us to identify robust PHA and SEB mode of action related phenotypic changes in PBMC immune cell lineages. Conclusively, we present a novel technique allowing the barcoded acquisition and analysis of PBMCs from up to 20 different donors and present a valid basis for the future development of complex immunophenotyping protocols.

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Section: Methodsmentioning

confidence: 99%

Barcoding of live peripheral blood mononuclear cells to assess immune cell phenotypes using full spectrum flow cytometry

Junker

Teixeira

2022

Cytometry Pt A

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“…Indeed, the percentage of positive cells is linked to some extent with the number of molecules per cell, but the exact relationship is unknown. 33 Hence, it was assumed that these RO calculations based on MFI and % PD-1(+) cells result in similar values.…”

Section: Methodsmentioning

confidence: 99%

Receptor occupancy assessment and interpretation in terms of quantitative systems pharmacology: nivolumab case study

Shchelokov¹,

Demin²

2022

mAbs

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Receptor occupancy assays applied in clinical studies provide insights into pharmacokinetic-pharmacodynamic relationships for therapeutic antibodies. When measured by different assays, however, receptor occupancy results can be controversial, as was observed for nivolumab, a monoclonal antibody targeting programmed cell death 1 (PD-1) receptor. We suggested an explanation of results obtained and a mechanistic approach based on specific features of the receptor occupancy assays: measurement of the free or bound receptor, normalized to the baseline or at each time point. The approach was evaluated against controversial clinical data on PD-1 receptor occupancy by nivolumab. It was shown that receptor occupancy measured by different assays might vary substantially if the internalization rate of the bound receptor is higher than the rate of degradation of the free receptor. Equations proposed in this work can be applied in quantitative systems pharmacology models to describe target receptor occupancy by different therapeutic antibodies.

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“…Details on the specific disease of the case study cannot be disclosed due to confidentiality reasons; however, the scientific investigation and simulations carried out are presented in a deidentified manner to provide useful insights for future applications. Although it was ultimately decided not to pursue this approach, the research conducted here provides a general framework that can be applied to any disease area where RO is expected to be related to drug efficacy, such as (but not limited to) monoclonal therapeutic antibodies in oncology (Junker et al, 2021;N. Jones et al, 2022), or some neurological disorders (Srinivas et al, 2018).…”

Section: Motivating Case Study In Immunoinflammationmentioning

confidence: 99%

Incorporation of healthy volunteers data on receptor occupancy into a phase II proof‐of‐concept trial using a Bayesian dynamic borrowing design

Di Stefano,

Rodrigues,

Galtier

et al. 2023

Biometrical J

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Receptor occupancy in targeted tissues measures the proportion of receptors occupied by a drug at equilibrium and is sometimes used as a surrogate of drug efficacy to inform dose selection in clinical trials. We propose to incorporate data on receptor occupancy from a phase I study in healthy volunteers into a phase II proof‐of‐concept study in patients, with the objective of using all the available evidence to make informed decisions. A minimal physiologically based pharmacokinetic modeling is used to model receptor occupancy in healthy volunteers and to predict it in the patients of a phase II proof‐of‐concept study, taking into account the variability of the population parameters and the specific differences arising from the pathological condition compared to healthy volunteers. Then, given an estimated relationship between receptor occupancy and the clinical endpoint, an informative prior distribution is derived for the clinical endpoint in both the treatment and control arms of the phase II study. These distributions are incorporated into a Bayesian dynamic borrowing design to supplement concurrent phase II trial data. A simulation study in immuno‐inflammation demonstrates that the proposed design increases the power of the study while maintaining a type I error at acceptable levels for realistic values of the clinical endpoint.

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A human receptor occupancy assay to measure anti‐PD‐1 binding in patients with prior anti‐PD‐1

Cited by 12 publications

References 52 publications

Barcoding of live peripheral blood mononuclear cells to assess immune cell phenotypes using full spectrum flow cytometry

Barcoding of live peripheral blood mononuclear cells to assess immune cell phenotypes using full spectrum flow cytometry

Receptor occupancy assessment and interpretation in terms of quantitative systems pharmacology: nivolumab case study

Incorporation of healthy volunteers data on receptor occupancy into a phase II proof‐of‐concept trial using a Bayesian dynamic borrowing design

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