Abstract 2073: Transcriptome profiles of cancer stem-like cells in patient-derived diffuse intrinsic pontine glioma (DIPG)
Authors: Galbán Stefanie , Espinoza Carlos , Bedi Karan , Maachani Uday B. , Souweidane Mark M. , Ljungman Mats , Dort Marcian Van , Ross Brian D. 
Published: August 2018

Diffuse intrinsic pontine glioma (DIPG) is a rare, but lethal childhood cancer with a 5-year survival less than 1 %. Genetic profiling of DIPG biopsies and post-mortem tissue have recently identified mutations in PI3KCA, PTEN, TP53, ATM/MPL, histones and PDGF receptor overexpression. PI3KCA and PTEN mutations as well as PDGF receptor overexpression indicate upregulation of the PI3K/AKT/mTOR signaling axis, representing druggable targets. We recently developed a multifunctional kinase inhibitor (ST-182), which targets the PI3K/AKT/mTOR and MAPK pathways which is often upregulated in various malignancies as a compensatory mechanism when PI3K is inhibited. We evaluated ST-182's efficacy for targeting these pathways in patient derived DIPG by western blotting and reverse phase protein array analysis (RPPA). Phosphorylation changes of ERK and AKT, downstream signaling inhibition as well as diminished proliferation was shown in two DIPG cell lines (SU-DIPGIV and XIII) when treated with ST-182, indicating efficacy of co-targeting these pathways as a new therapeutic advance for DIPG.

FACS analysis of DIPG cells (SU-DIPGXIII) identified a large percentage (&gt;10%) of DIPG cells as ALDH positive indicating aggressive stem like features. Characterization of these distinct DIPG populations (ALDH+,-) at the transcriptome level was performed to understand differences in pathway signaling and to identify potential drug resistance mechanisms to ST-182. Utilizing an innovative transcriptome analysis approach, we identified elevated levels of MYC, E2F and DNA repair genes in ALDH+ cells, supporting stem like phenotype of ALDH+ DIPG cells. MYC has long been identified as a crucial player in maintaining embryonic stem cell pluripotency and self-renewal, whereas E2F provide transcriptional control of stem cell fate and DNA repair mechanisms maintain and regulate cancer stem cells. Pharmacological targeting of MAPK/PI3K/mTOR by ST-182 demonstrated up regulation of NFkB, apoptosis, hypoxia, p53 and inflammatory response in ALDH+ and ALDH- cells and down-regulation of MYC, E2F and DNA replication indicating efficacy of targeting these pathways in preventing/reversing stem-like phenotypes in the ALDH+ cell population.

Our findings indicate efficacy of ST-182 for the treatment of ALDH+ cancer stem cells providing impetus for evaluation of molecularly targeted MAPK/PI3K/mTOR therapy. Our comprehensive transcriptome studies provide a new direction for the treatment of DIPG through novel insights into the underlying transcriptomic basis of drug resistant cancer stem cells. Development of new compounds such as ST-182 provides opportunities to implement precision medicine to improve treatment outcomes for DIPG patients.

Citation Format: Stefanie Galbán, Carlos Espinoza, Karan Bedi, Uday B. Maachani, Mark M. Souweidane, Mats Ljungman, Marcian Van Dort, Brian D. Ross. Transcriptome profiles of cancer stem-like cells in patient-derived diffuse intrinsic pontine glioma (DIPG) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2073.