Feasibility study of using high‐throughput drug sensitivity testing to target recurrent glioblastoma stem cells for individualized treatment

Abstract

BackgroundDespite the well described heterogeneity in glioblastoma (GBM), treatment is standardized, and clinical trials investigate treatment effects at population level. Genomics‐driven oncology for stratified treatments allow clinical decision making in only a small minority of screened patients. Addressing tumor heterogeneity, we aimed to establish a clinical translational protocol in recurrent GBM (recGBM) utilizing autologous glioblastoma stem cell (GSC) cultures and automated high‐throughput drug sensitivity and resistance testing (DSRT) for individualized treatment within the time available for clinical application. ResultsFrom ten patients undergoing surgery for recGBM, we established individual cell cultures and characterized the GSCs by functional assays. 7/10 GSC cultures could be serially expanded. The individual GSCs displayed intertumoral differences in their proliferative capacity, expression of stem cell markers and variation in their in vitro and in vivo morphology. We defined a time frame of 10 weeks from surgery to complete the entire pre‐clinical work‐up; establish individualized GSC cultures, evaluate drug sensitivity patterns of 525 anticancer drugs, and identify options for individualized treatment. Within the time frame for clinical translation 5/7 cultures reached sufficient cell yield for complete drug screening. The DSRT revealed significant intertumoral heterogeneity to anticancer drugs (p < 0.0001). Using curated reference databases of drug sensitivity in GBM and healthy bone marrow cells, we identified individualized treatment options in all patients. Individualized treatment options could be selected from FDA‐approved drugs from a variety of different drug classes in all cases. ConclusionsIn recGBM, GSC cultures could successfully be established in the majority of patients. The individual cultures displayed intertumoral heterogeneity in their in vitro and in vivo behavior. Within a time frame for clinical application, we could perform DSRT in 50% of recGBM patients. The DSRT revealed a remarkable intertumoral heterogeneity in sensitivity to anticancer drugs in recGBM that could allow tailored therapeutic options for functional precision medicine.

Double Recombinant Vaccinia Virus: A Candidate Drug against Human Glioblastoma

“…Challenges exist regarding the clinical application of OVs, such as the resistance of glioblastoma, immunosuppressive microenvironments, or the presence of the bloodbrain barrier [18,19]. There also is complicated internal heterogeneity of glioblastoma at the cellular and molecular level [20] as well as with the selection of relevant models for preclinical research [10]. One of the common preclinical models for studying the biology of glioblastoma and its response to therapy are cell cultures obtained from patient tumor samples and cultured under conditions of neurosphere formation [21].…”

The important role of routine cytopathology in pediatric precision oncology

“…10,11 It has been proven that HTS using primary cultures accurately predicts in vivo efficacy in animal models. 7,8,12 Therefore, conducting HTS using primary cultures with a library of approved drugs, integrated with molecular profiling, provides a clinically relevant approach to tailoring treatment options for patients with cancer. 13 In solid tumor oncology, the relative paucity and impaired viability of tumor cells limit the capacity of using in vitro drug sensitivity data to influence patient management in a clinically relevant time frame.…”

“…Preclinical models, including in vivo and in vitro models, such as primary cultures, syngeneic mouse models, and patient‐derived xenografts (PDXs), are widely used to validate drug efficacy and identify additional therapeutic options for patients with cancer 7,8 Although PDX models more accurately recapitulate the genetic characteristics and heterogeneity of the original tumor, 6 the practicality of using drug efficacy data obtained from PDXs to guide patient management in a clinically relevant time frame is unclear 9 . Primary cultures, derived from freshly dissociated patient tumors or PDXs, are a valid tool for preclinical analyses 10,11 .…”

“…Primary cultures, derived from freshly dissociated patient tumors or PDXs, are a valid tool for preclinical analyses 10,11 . It has been proven that HTS using primary cultures accurately predicts in vivo efficacy in animal models 7,8,12 . Therefore, conducting HTS using primary cultures with a library of approved drugs, integrated with molecular profiling, provides a clinically relevant approach to tailoring treatment options for patients with cancer 13 …”

Robot technology identifies a Parkinsonian therapeutics repurpose to target stem cells of glioblastoma

“…From a clinical-translational prospective, we think that this technology would be useful to determine personal chemo-responsome of primary cells freshly derived from surgical resection specimens. Our robotics complete the list of liquid handling devices to do drug testing on tumors in vitro [ 2 , 17 ], thereby allowing this approach to be implementable in different lab settings and suitable for a variety of disease types and disease modeling strategies. We argue that our system will also be useful for testing drug resistance on more complex cell model culture systems that require pre-coating of culture plates – such as human induced pluripotent stem cells – mainly due to the detailed and programmable pipetting basis of liquid transfer in our device, which emulate manual pipetting on a high scale and cannot be facilitated by printing-based drug screening assays [ 18 ].…”