Image-based ex-vivo drug screening for patients with aggressive haematological malignancies: interim results from a single-arm, open-label, pilot study

Abstract

SummaryBackgroundPatients with refractory or relapsed haematological malignancies have few treatment options and short survival times. Identification of effective therapies with genomic-based precision medicine is hampered by intratumour heterogeneity and incomplete understanding of the contribution of various mutations within specific cancer phenotypes. Ex-vivo drug-response profiling in patient biopsies might aid effective treatment identification; however, proof of its clinical utility is limited.MethodsWe investigated the feasibility and clinical impact of multiparametric, single-cell, drug-response profiling in patient biopsies by immunofluorescence, automated microscopy, and image analysis, an approach we call pharmacoscopy. First, the ability of pharmacoscopy to separate responders from non-responders was evaluated retrospectively for a cohort of 20 newly diagnosed and previously untreated patients with acute myeloid leukaemia. Next, 48 patients with aggressive haematological malignancies were prospectively evaluated for pharmacoscopy-guided treatment, of whom 17 could receive the treatment. The primary endpoint was progression-free survival in pharmacoscopy-treated patients, as compared with their own progression-free survival for the most recent regimen on which they had progressive disease. This trial is ongoing and registered with ClinicalTrials.gov, number NCT03096821.FindingsPharmacoscopy retrospectively predicted the clinical response of 20 acute myeloid leukaemia patients to initial therapy with 88·1% accuracy. In this interim analysis, 15 (88%) of 17 patients receiving pharmacoscopy-guided treatment had an overall response compared with four (24%) of 17 patients with their most recent regimen (odds ratio 24·38 [95% CI 3·99–125·4], p=0·0013). 12 (71%) of 17 patients had a progression-free survival ratio of 1·3 or higher, and median progression-free survival increased by four times, from 5·7 (95% CI 4·1–12·1) weeks to 22·6 (7·4–34·0) weeks (hazard ratio 3·14 [95% CI 1·37–7·22], p=0·0075).InterpretationRoutine clinical integration of pharmacoscopy for treatment selection is technically feasible, and led to improved treatment of patients with aggressive refractory haematological malignancies in an initial patient cohort, warranting further investigation.FundingAustrian Academy of Sciences; European Research Council; Austrian Science Fund; Austrian Federal Ministry of Science, Research and Economy; National Foundation for Research, Technology and Development; Anniversary Fund of the Austrian National Bank; MPN Research Foundation; European Molecular Biology Organization; and Swiss National Science Foundation.

Precision Medicine in Oncology: In Vitro Drug Sensitivity and Resistance Test (DSRT) for Selection of Personalized Anticancer Therapy

“…Cell viability can be also quantified by staining cells followed by microscopy . Hoechst 33 342 and 4′,6‐diamidino‐2‐phenylindole (DAPI) are often used to stain cell nuclei and estimate total cell number . There are a number of viability dyes available including CyQuant, Calcein AM, and propidium iodide that can distinguish between live and dead cells.…”

“…In addition, using microscopy‐based read‐out opens possibilities for more in‐depth analysis of treated cell population. For example, performing immunostaining with antibodies against specific marker of cancer cells enables distinguishing between response of cancer and healthy cells, identifying if drug is specific or generally toxic …”

“…Snijder et al. developed a concept of pharmacoscopy, which is a methodology for ex vivo drug response profiling . This methodology is based on immunofluorescence, automated microscopy, and image analysis of biopsies cells introduced to anticancer drugs in vitro.…”

“…In addition, this methodology was used to profile 48 patients with aggressive hematological malignancies, 17 of whom received the pharmacoscopy‐guided treatment resulted in partial and complete remissions. Pharmacoscopy is evaluated in ongoing clinical trial at the moment …”

The Convergence of Stem Cell Technologies and Phenotypic Drug Discovery

“…Tracking datasets for compound libraries across research institutions and correlating phenotypes acquired by different individuals under different conditions could help turning the vision of in silico modeling into a reality by distilling core multidimensional phenotypic changes out of a heterogeneous biological sample space. While we are not aware of such advances in the stem cell field, innovative approaches including patient-specific, high-content immunofluorescence-driven approaches in complex co-culture formats with a cross-patient longitudinal integration of datasets exists in clinical cancer research, in a technique called pharmacoscopy (Snijder et al, 2017). With the growing availability of stem cell banks coupled with in-depth patient records, applying such comprehensive approaches applied to stem cell screens and compound libraries will drive additional insight into health, developmental, and disease aspects across cell lines and patients, thus further broadening the insight and predictability of phenotypic changes in stem cell-based screens.…”