Real-time cell viability monitoring for high-throughput drug screening using tumor xenograft-derived cells

Elham Esmaeilishirazifard; Daniel Guerrero-Romero; Allan J.W. Lui; Abigail Shea; Long V. Nguyen; Maurizio Callari; Riccardo Masina; Kevin Tu; Richard Baird; Paul Edwards; Wendy Greenwood; Steve Fuller; Claire Crafter; Mandy Lawson; Alejandra Bruna; Oscar M. Rueda; Carlos Caldas

doi:10.1016/j.crmeth.2026.101464

Real-time cell viability monitoring for high-throughput drug screening using tumor xenograft-derived cells

Elham Esmaeilishirazifard
, Daniel Guerrero-Romero
, Allan J.W. Lui
, Abigail Shea
, Long V. Nguyen
, Maurizio Callari
, Riccardo Masina
, Kevin Tu
, Richard Baird
, Paul Edwards
, Wendy Greenwood
, Steve Fuller
, Claire Crafter
, Mandy Lawson
, Alejandra Bruna
, Oscar M. Rueda^*
, Carlos Caldas^*

^*Corresponding author for this work

Department of Immunology and Cancer Research

Research output: Contribution to journal › Article › peer-review

Abstract

Patient-derived tumor xenografts (PDTXs) recapitulate the molecular and phenotypic heterogeneity of human cancers, making them valuable pre-clinical models for cancer drug development. However, high-throughput drug screening (HTDS) using ex vivo short-term cultures of PDTX-derived tumor cells (PDTCs) is hindered by endpoint viability assays that provide only static measures of drug response. Here, we establish an optimized a screening platform by validating the RealTime-Glo (RTG) bioluminescent assay for dynamic, real-time measurements of PDTC viability. We further introduce an analytical metric to quantify drug responses independent of cell growth rate. Using this approach, we screened 67 compounds across 43 breast cancer PDTCs and revealed model-specific pharmacodynamic heterogeneity. Our PDTC-based HTDS pipeline improves assay robustness and offers an enhanced platform for leveraging patient-derived xenograft models in precision medicine.

Original language	English
Article number	101464
Journal	Cell Reports Methods
Volume	6
Issue number	7
DOIs	https://doi.org/10.1016/j.crmeth.2026.101464
State	Published - 20 Jul 2026

Bibliographical note

Publisher Copyright:
© 2026

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

CP: cancer biology
PDTCs
PDTX-derived tumor cells
PDTXs
breast cancer
drug response kinetics
dynamic viability monitoring
ex vivo drug sensitivity
high-throughput drug screening
isotonic regression modeling
patient-derived tumor xenografts
pharmacodynamic profiling
real-time viability monitoring

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10.1016/j.crmeth.2026.101464

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Esmaeilishirazifard, E., Guerrero-Romero, D., Lui, A. J. W., Shea, A., Nguyen, L. V., Callari, M., Masina, R., Tu, K., Baird, R., Edwards, P., Greenwood, W., Fuller, S., Crafter, C., Lawson, M., Bruna, A., Rueda, O. M., & Caldas, C. (2026). Real-time cell viability monitoring for high-throughput drug screening using tumor xenograft-derived cells. Cell Reports Methods, 6(7), Article 101464. https://doi.org/10.1016/j.crmeth.2026.101464

@article{746f0a813d3d461dadc36698e1601294,

title = "Real-time cell viability monitoring for high-throughput drug screening using tumor xenograft-derived cells",

abstract = "Patient-derived tumor xenografts (PDTXs) recapitulate the molecular and phenotypic heterogeneity of human cancers, making them valuable pre-clinical models for cancer drug development. However, high-throughput drug screening (HTDS) using ex vivo short-term cultures of PDTX-derived tumor cells (PDTCs) is hindered by endpoint viability assays that provide only static measures of drug response. Here, we establish an optimized a screening platform by validating the RealTime-Glo (RTG) bioluminescent assay for dynamic, real-time measurements of PDTC viability. We further introduce an analytical metric to quantify drug responses independent of cell growth rate. Using this approach, we screened 67 compounds across 43 breast cancer PDTCs and revealed model-specific pharmacodynamic heterogeneity. Our PDTC-based HTDS pipeline improves assay robustness and offers an enhanced platform for leveraging patient-derived xenograft models in precision medicine.",

keywords = "CP: cancer biology, PDTCs, PDTX-derived tumor cells, PDTXs, breast cancer, drug response kinetics, dynamic viability monitoring, ex vivo drug sensitivity, high-throughput drug screening, isotonic regression modeling, patient-derived tumor xenografts, pharmacodynamic profiling, real-time viability monitoring",

author = "Elham Esmaeilishirazifard and Daniel Guerrero-Romero and Lui, \{Allan J.W.\} and Abigail Shea and Nguyen, \{Long V.\} and Maurizio Callari and Riccardo Masina and Kevin Tu and Richard Baird and Paul Edwards and Wendy Greenwood and Steve Fuller and Claire Crafter and Mandy Lawson and Alejandra Bruna and Rueda, \{Oscar M.\} and Carlos Caldas",

note = "Publisher Copyright: {\textcopyright} 2026",

year = "2026",

month = jul,

day = "20",

doi = "10.1016/j.crmeth.2026.101464",

language = "אנגלית",

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Esmaeilishirazifard, E, Guerrero-Romero, D, Lui, AJW, Shea, A, Nguyen, LV, Callari, M, Masina, R, Tu, K, Baird, R, Edwards, P, Greenwood, W, Fuller, S, Crafter, C, Lawson, M, Bruna, A, Rueda, OM & Caldas, C 2026, 'Real-time cell viability monitoring for high-throughput drug screening using tumor xenograft-derived cells', Cell Reports Methods, vol. 6, no. 7, 101464. https://doi.org/10.1016/j.crmeth.2026.101464

TY - JOUR

T1 - Real-time cell viability monitoring for high-throughput drug screening using tumor xenograft-derived cells

AU - Esmaeilishirazifard, Elham

AU - Guerrero-Romero, Daniel

AU - Lui, Allan J.W.

AU - Shea, Abigail

AU - Nguyen, Long V.

AU - Callari, Maurizio

AU - Masina, Riccardo

AU - Tu, Kevin

AU - Baird, Richard

AU - Edwards, Paul

AU - Greenwood, Wendy

AU - Fuller, Steve

AU - Crafter, Claire

AU - Lawson, Mandy

AU - Bruna, Alejandra

AU - Rueda, Oscar M.

AU - Caldas, Carlos

PY - 2026/7/20

Y1 - 2026/7/20

N2 - Patient-derived tumor xenografts (PDTXs) recapitulate the molecular and phenotypic heterogeneity of human cancers, making them valuable pre-clinical models for cancer drug development. However, high-throughput drug screening (HTDS) using ex vivo short-term cultures of PDTX-derived tumor cells (PDTCs) is hindered by endpoint viability assays that provide only static measures of drug response. Here, we establish an optimized a screening platform by validating the RealTime-Glo (RTG) bioluminescent assay for dynamic, real-time measurements of PDTC viability. We further introduce an analytical metric to quantify drug responses independent of cell growth rate. Using this approach, we screened 67 compounds across 43 breast cancer PDTCs and revealed model-specific pharmacodynamic heterogeneity. Our PDTC-based HTDS pipeline improves assay robustness and offers an enhanced platform for leveraging patient-derived xenograft models in precision medicine.

AB - Patient-derived tumor xenografts (PDTXs) recapitulate the molecular and phenotypic heterogeneity of human cancers, making them valuable pre-clinical models for cancer drug development. However, high-throughput drug screening (HTDS) using ex vivo short-term cultures of PDTX-derived tumor cells (PDTCs) is hindered by endpoint viability assays that provide only static measures of drug response. Here, we establish an optimized a screening platform by validating the RealTime-Glo (RTG) bioluminescent assay for dynamic, real-time measurements of PDTC viability. We further introduce an analytical metric to quantify drug responses independent of cell growth rate. Using this approach, we screened 67 compounds across 43 breast cancer PDTCs and revealed model-specific pharmacodynamic heterogeneity. Our PDTC-based HTDS pipeline improves assay robustness and offers an enhanced platform for leveraging patient-derived xenograft models in precision medicine.

KW - CP: cancer biology

KW - PDTCs

KW - PDTX-derived tumor cells

KW - PDTXs

KW - breast cancer

KW - drug response kinetics

KW - dynamic viability monitoring

KW - ex vivo drug sensitivity

KW - high-throughput drug screening

KW - isotonic regression modeling

KW - patient-derived tumor xenografts

KW - pharmacodynamic profiling

KW - real-time viability monitoring

UR - https://www.scopus.com/pages/publications/105040116808

U2 - 10.1016/j.crmeth.2026.101464

DO - 10.1016/j.crmeth.2026.101464

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C2 - 42202786

AN - SCOPUS:105040116808

SN - 2667-2375

VL - 6

JO - Cell Reports Methods

JF - Cell Reports Methods

IS - 7

M1 - 101464

ER -

Real-time cell viability monitoring for high-throughput drug screening using tumor xenograft-derived cells

Abstract

Bibliographical note

UN SDGs

Keywords

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