PTEN Regulates PI(3,4)P2 Signaling Downstream of Class I PI3K

Mouhannad Malek,Véronique Juvin,Izabella Niewczas,Satoshi Eguchi,Pınar Pir,Hiroki Nakanishi,Dominik Spensberger,Atsushi Koizumi,Marine Imbert,Karen E Anderson,Phillip T Hawkins,Alexander Gray,David Barneda,Anna Kielkowska,Len R Stephens,Takehiko Sasaki,Nicolas Le Novère,Tomonori Habuchi,Junko Sasaki,Jonathan Clark,Alexandre Valayer,Tamara Chessa,Sabina C Cosulich ,Sören Beinke ,Sérgio Luís Felisbino ,Vladimir Yu Kiselev

doi:10.1016/j.molcel.2017.09.024

Abstract

SummaryThe PI3K signaling pathway regulates cell growth and movement and is heavily mutated in cancer. Class I PI3Ks synthesize the lipid messenger PI(3,4,5)P3. PI(3,4,5)P3 can be dephosphorylated by 3- or 5-phosphatases, the latter producing PI(3,4)P2. The PTEN tumor suppressor is thought to function primarily as a PI(3,4,5)P3 3-phosphatase, limiting activation of this pathway. Here we show that PTEN also functions as a PI(3,4)P2 3-phosphatase, both in vitro and in vivo. PTEN is a major PI(3,4)P2 phosphatase in Mcf10a cytosol, and loss of PTEN and INPP4B, a known PI(3,4)P2 4-phosphatase, leads to synergistic accumulation of PI(3,4)P2, which correlated with increased invadopodia in epidermal growth factor (EGF)-stimulated cells. PTEN deletion increased PI(3,4)P2 levels in a mouse model of prostate cancer, and it inversely correlated with PI(3,4)P2 levels across several EGF-stimulated prostate and breast cancer lines. These results point to a role for PI(3,4)P2 in the phenotype caused by loss-of-function mutations or deletions in PTEN.

Highlights

The class I PI3K-signaling pathway is part of the large regulatory network that allows various cell surface receptors to control cell function (Hawkins and Stephens, 2015; Vanhaesebroeck et al, 2010)
A Method to Measure PI(3,4)P2 and PI(4,5)P2 by high-performance liquid chromatography (HPLC)-MS We have previously shown that methylation of the acidic phosphate groups of polyphosphoinositides with trimethylsilyl (TMS)-diazomethane allows sensitive detection of these molecules by HPLC-electrospray ionization-mass spectrometry (HPLC-ESI-MS) (Kielkowska et al, 2014)
SiRNA knockdown of SHIP2 in PTEN-KO cells or CRISPR/Cas9-mediated deletion of SHIP2 expression in PTEN-KO cells resulted in a substantial, synergistic elevation in peak PI(3,4,5)P3 levels (3- to 4-fold), though in each case the PI(3,4,5)P3 response remained transient, with levels falling within 5–15 min of epidermal growth factor (EGF) stimulation (Figure 2A). These results clearly identify PTEN and SHIP2 as phosphatases that act on PI(3,4,5)P3 during EGF stimulation, but they suggest that each can substantially compensate for loss of the other

Summary

Introduction

The class I PI3K-signaling pathway is part of the large regulatory network that allows various cell surface receptors to control cell function (Hawkins and Stephens, 2015; Vanhaesebroeck et al, 2010) This pathway plays a important role in the mechanisms that allow growth factor receptors to regulate cell growth. The best studied of these effectors are the PH domain-containing serine/threonine protein kinases PDK-1 and AKT1/2, which indirectly regulate the mTORC1 complex and promote anabolic growth and survival (Dibble and Cantley, 2015; Engelman et al, 2006) This pathway is heavily mutated in human cancers, harboring several prevalent oncogenes, including genes encoding PI3K subunits and AKT, and several tumor suppressors, for example, PTEN and INPP4B (Fruman and Rommel, 2014; Mayer and Arteaga, 2016; Okkenhaug et al, 2016; Thorpe et al, 2015)

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