Abstract Background: Activation of the PI3K pathway is implicated in a number of tumour types, including haematological malignancies. Agents that target this pathway at different levels (e.g. RTK, PI3K, mTOR etc) have shown promising activity in preclinical models and in early phase trials. We have therefore investigated the activity of novel, potent, pan- and isoform-selective PI3K pathway inhibitors in lymphoma and myeloma models. Methods: Compounds studied included those with selectivity for mTOR (INK128, mTOR IC50 1.6 nM), PI3Kδ/γ (INK713, INK1048, IC50s <10 nM for δ/γ isoforms, >50 nM for others) and PI3Kδ/γ/β (INK1138, IC50s <10 nM for each isoform). IC87114 (δ- selective), GDC-0941 (α/δ selective), rapamycin and doxorubicin were included as comparators. Agents were studied in a panel of lymphoma and myeloma cell lines, in normal PBMCs and in primary tumours cultured with stromal cells. Effects on cell viability and proliferation were assessed using the Guava Viacount assay, on apoptosis induction by annexin V labelling, and on cell cycle distribution by flow cytometry. PI3K pathway activity and inhibition was determined by Western blotting as well as mass spectrometry. Results: In cell lines, inhibition of mTOR by INK128 resulted in potent inhibition of cell proliferation, in many at concentrations as low as 10 nM. Selective PI3K inhibitors were less potent anti-proliferative agents, although activity was seen in cells with activation of the PI3K pathway as determined by western blot analysis. For all compounds tested, the major impact was on cell proliferation rather than cell viability. PI3K inhibitors showed little effect on cell viability at concentrations < 1µM. The effect of these compounds in primary mantle cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia and myeloma samples was variable, both between and within tumour types, possibly due to differences in basal PI3K pathway activity. Effects in normal PBMCs were markedly different to those seen in tumour cells, with a concentration-dependent increase in cell number with either mTOR or PI3K inhibition. Conclusion: Selective PI3K pathway inhibition, particularly at the level of mTOR, results in cytostatic, and at higher concentrations cytotoxic, responses in haematological malignancies. The variable activity of these compounds between cell lines and primary samples suggests it will be important to characterise the activity of the PI3K pathway in individual tumours to identify patients likely to benefit from such agents. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4489.