Metabolic biomarkers for response to PI3K inhibition in basal-like breast cancer

Abstract

IntroductionThe phosphatidylinositol 3-kinase (PI3K) pathway is frequently activated in cancer cells through numerous mutations and epigenetic changes. The recent development of inhibitors targeting different components of the PI3K pathway may represent a valuable treatment alternative. However, predicting efficacy of these drugs is challenging, and methods for therapy monitoring are needed. Basal-like breast cancer (BLBC) is an aggressive breast cancer subtype, frequently associated with PI3K pathway activation. The objectives of this study were to quantify the PI3K pathway activity in tissue sections from xenografts representing basal-like and luminal-like breast cancer before and immediately after treatment with PI3K inhibitors, and to identify metabolic biomarkers for treatment response.MethodsTumor-bearing animals (n = 8 per treatment group) received MK-2206 (120 mg/kg/day) or BEZ235 (50 mg/kg/day) for 3 days. Activity in the PI3K/Akt/mammalian target of rapamycin pathway in xenografts and human biopsies was evaluated using a novel method for semiquantitative assessment of Aktser473 phosphorylation. Metabolic changes were assessed by ex vivo high-resolution magic angle spinning magnetic resonance spectroscopy.ResultsUsing a novel dual near-infrared immunofluorescent imaging method, basal-like xenografts had a 4.5-fold higher baseline level of pAktser473 than luminal-like xenografts. Following treatment, basal-like xenografts demonstrated reduced levels of pAktser473 and decreased proliferation. This correlated with metabolic changes, as both MK-2206 and BEZ235 reduced lactate concentration and increased phosphocholine concentration in the basal-like tumors. BEZ235 also caused increased glucose and glycerophosphocholine concentrations. No response to treatment or change in metabolic profile was seen in luminal-like xenografts. Analyzing tumor sections from five patients with BLBC demonstrated that two of these patients had an elevated pAktser473 level.ConclusionThe activity of the PI3K pathway can be determined in tissue sections by quantitative imaging using an antibody towards pAktser473. Long-term treatment with MK-2206 or BEZ235 resulted in significant growth inhibition in basal-like, but not luminal-like, xenografts. This indicates that PI3K inhibitors may have selective efficacy in basal-like breast cancer with increased PI3K signaling, and identifies lactate, phosphocholine and glycerophosphocholine as potential metabolic biomarkers for early therapy monitoring. In human biopsies, variable pAktser473 levels were observed, suggesting heterogeneous PI3K signaling activity in BLBC.

Treatment with aromatase inhibitors stimulates the expression of epidermal growth factor receptor-1 and neuregulin 1 in ER positive/HER-2/neu non-amplified primary breast cancers

“…Negative control sections were prepared by incubating only with the secondary antibody. Near infrared immunofluorescent imaging was performed as described previously [32]. The pAkt Ser473 staining intensity was measured using Image Studio 3.1.…”

Probing the PI3K/Akt/mTor pathway using 31P-NMR spectroscopy: routes to glycogen synthase kinase 3

“…Several in-vitro and in-vivo studies1724252627 have applied 31 P-NMR to measure phospholipid metabolite levels in tumour/cells responding to drugs targeting the PI3K/Akt/mTOR but the findings have not demonstrated a clear consensus especially with respect to their effects on PCho content. Since metabolites may serve as prognostic or predictive biomarkers, we aimed to explore the effect of drugs inhibiting several components of PI3K signalling (see Fig.…”

Proteomic Characterization of Breast Cancer Xenografts Identifies Early and Late Bevacizumab-Induced Responses and Predicts Effective Drug Combinations

“…The lack of effect from Iressa monotherapy in the in vivo experiments may be explained by the low baseline level of PTEN expression in the basal-like, compared to the luminal-like, model [30], as PTEN loss has been shown to confer resistance to EGFR kinase inhibitors [31]. Another explanation for the lack of Iressa effect may the targeting of an upstream growth factor receptor, which may not be sufficient to inhibit tumor growth due to the redundancy in downstream pathway activators.…”