Avalon Pharmaceuticals, Inc.

To give a state-of-the-art overview on the promise of three-dimensional (3-D) culture systems for anticancer drug development, with particular emphasis on multicellular tumor spheroids (MCTS).

Cell-based assays have become an integral component in many stages of routine anti-tumor drug testing. However, they are almost always based on homogenous monolayer or suspension cultures and thus represent a rather artificial cellular environment. 3-D cultures--such as the well established spheroid culture system--better reflect the in vivo behavior of cells in tumor tissues and are increasingly recognized as valuable advanced tools for evaluating the efficacy of therapeutic intervention. The present article summarizes past and current applications and particularly discusses technological challenges, required improvements and recent progress with the use of the spheroid model in experimental therapeutics, as a basis for sophisticated drug/therapy screening. A brief overview is given focusing on the nomenclature of spherical 3-D cultures, their potential to mimic many aspects of the pathophysiological situation in tumors, and currently available protocols for culturing and analysis. A list of spheroid-forming epithelial cancer cell lines of different origin is provided and the recent trend to use spheroids for testing combination treatment strategies is highlighted. Finally, various spheroid co-culture approaches are presented that have been established to study heterologous cell interactions in solid tumors and thereby are able to reflect the cellular tumor environment with increasing accuracy. The intriguing observation that in order to retain certain tumor initiating cell properties, some primary tumor cell populations must be maintained exclusively in 3-D culture is mentioned, adding a new but fascinating challenge for future therapeutic campaigns.

The long arm of chromosome 6 is frequently deleted in diverse human neoplasms. Our previous study showed a minimum deletion region between markers D6S1056 and D6S300 on chromosome 6q in primary prostate cancer (CaP). In this study, we further refined a 200-kb minimal region of deletion (6qTSG1) centered around D6S1013 marker. The 6qTSG1 transcripts contained complex multiple splicing variants with low or absent expression in CaP cells. None of the transcripts identified contained open reading frames that code for a protein in the NCBI database. The expression of 6qTSG transcripts revealed interesting hormonal regulation relevant to CaP biology. Expression of 6q TSG transcript was induced in LNCaP cells that were cultured in charcoal-stripped serum medium suggesting an upregulation of 6qTSG transcript by androgen ablation and cell growth inhibition/apoptosis. Induction of 6qTSG1 expression in response to androgen ablation was abrogated in androgen-independent derivatives of LNCaP cells. In summary, we have defined a candidate CaP suppressor locus on chromosome 6q16.1, and deletions of this locus are frequently associated with prostate tumorigenesis. In the light of emerging role of noncoding RNAs in cancer biology including CaP, future investigations of 6qTSG11 locus is warranted.

Progress in cancer treatment has been slow, and the outlook for curing cancer is only marginally different from the situation a decade ago. Paradoxically, although the pharmaceutical industry has stepped up costly discovery research and drug development, approvals are on the decline and pipelines are dwindling. In an effort to reduce the number of drug failures and curtail burgeoning R&D costs, drug companies are exploring the use of biomarkers to evaluate toxicity and efficacy earlier in the development process. Biomarkers hold promise for optimization in dosing, adverse event prediction, efficacy evaluation, lead prioritization, and mechanism-of-action profiling of drug candidates. Furthermore, clinicians can use biomarkers to monitor patient response in clinical trials. In this perspective article, the authors explore the applications of cancer-related expression biomarkers in drug discovery and discuss how this will impact the industry and benefit the patient.