Feasibility and barriers to rapid establishment of patient-derived primary osteosarcoma cell lines in clinical management

Abstract

Osteosarcoma is a highly aggressive primary bone tumor that has seen little improvement in survival rates in the past three decades. Preclinical studies are conducted on a small pool of commercial cell lines which may not fully reflect the genetic heterogeneity of this complex cancer, potentially hindering translatability of in vitro results. Developing a single-site laboratory protocol to rapidly establish patient-derived primary cancer cell lines (PCCL) within a clinically actionable time frame of a few weeks will have significant scientific and clinical ramifications. These PCCL can widen the pool of available cell lines for study while patient-specific data could derive therapeutic correlation. This endeavor is exceedingly challenging considering the proposed time constraints. By proposing key definitions and a clear theoretical framework, this evaluation of osteosarcoma cell line establishment methodology over the past three decades assesses feasibility by identifying barriers and suggesting solutions, thereby facilitating systematic experimentation and optimization.

Keywords: Cancer; Technical aspects of cell biology.

Graphical abstract

Figure 1

Timeline of OS management with potential six-week time frame for PCCL establishment Standard multi-agent MAP chemotherapy regimen at our institution. All patients undergo a bone biopsy for histological subtyping as part of the diagnostic workup. Two cycles of NAC are given for a total of 10 weeks. Surgery is performed to remove the primary tumor. Four cycles of adjuvant chemotherapy commence postoperatively. There is a potential window of six-weeks between bone biopsy and the end of cycle 1, for PCCL to be established and tested. Adapted from Choong (2021). Abbreviations: Osteosarcoma (OS), primary cancer cell line (PCCL), neoadjuvant chemotherapy (NAC), high-dose MAP (methotrexate, doxorubicin, cisplatin), bx (biopsy), dx (diagnosis).

Figure 2

PRISMA Diagram outlining search strategy and eligible studies This diagram presents the flow of information through the systematic review process, following the PRISMA guidelines. The diagram outlines the number of records identified through database searching, additional records identified through other sources, records screened, records excluded, full-text articles assessed for eligibility, and the final number of articles included in the systematic review. Arrows indicate the flow of articles at each stage, with reasons for exclusion provided where applicable. The PRISMA flow diagram provides transparency and clarity regarding the selection process for inclusion of studies in the systematic review. Abbreviations: osteosarcoma (OS), patient-derived xenograft (PDX).

Figure 3

Workflow for establishing OS PCCL within clinical management This workflow summarizes the extracted data, barriers to feasibility, and potential solutions for establishment of PCCL within six weeks. The overall methodology is categorized into five Entry steps. The second column summarizes the extracted data and identifies barriers in each Entry. The final column suggests potential solutions to overcome said barriers, which serves as a useful guide for future experiments. Abbreviations: Osteosarcoma (OS), primary cancer cell lines (PCCL), extracellular matrix (ECM), International Cell Line Authentication Committee (ICLAC), neoadjuvant chemotherapy (NAC), short-tandem repeat (STR), next generation sequencing (NGS), real-time PCR (RT-PCR), immunohistochemistry (IHC), western blot (WB).