Feature Review

Synergy in activating class I PI3Ks

John E. Burke1 and Roger L. Williams2
1
Department of Biochemistry and Microbiology, University of Victoria, 3800 Finnerty Drive, Victoria BC, V8P 5C2, Canada
2
Medical Research Council (MRC) Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus,
Cambridge, CB2 0QH, UK

The class I phosphoinositide 3-kinases (PI3Ks) are lipid including growth, cell motility, cytoskeletal reorganiza-
kinases that transduce a host of cellular signals and tion, protein synthesis, cell metabolism, and immune cell
regulate a broad range of essential functions including differentiation (Figure 1B).
growth, proliferation, and migration. As such, PI3Ks The ability of class I PI3Ks to perform such a diverse set
have pivotal roles in diseases such as cancer, diabetes, of signaling roles in many tissues and systems requires
primary immune disorders, and inflammation. These tight regulation of their lipid kinase activity, controlled
enzymes are activated downstream of numerous acti- through the coordination of multiple upstream signaling
vating stimuli including receptor tyrosine kinases, G inputs. Key to the regulation of class I PI3Ks downstream
protein-coupled receptors (GPCRs), and the Ras super- of signaling inputs is the presence of regulatory subunits.
family of small G proteins. A major challenge is to deci- The class IA PI3Ks are composed of three different cata-
pher how each PI3K isoform is able to successfully lytic subunits (p110a, p110b, and p110d) which tightly
synergize these inputs into their intended signaling interact with one of five different regulatory subunits
function. This article highlights recent progress in char- (p85a, p85b, p55a, p50a, and p55g). The class IB PI3Ks
acterizing the molecular mechanisms of PI3K isoform- are composed of a single type of catalytic subunit (p110g)
specific activation pathways, as well as novel roles for which can interact with one of two different regulatory
PI3Ks in human diseases, specifically cancer and im- subunits (p84/p87 or p101). Both the catalytic and regula-
mune diseases. tory subunits of class IA PI3Ks are large multidomain
proteins, and structural and biochemical studies have
Class I PI3Ks are key signaling effectors defined their domain architecture and their mechanism
Phosphoinositide lipids are minor components of the plas- of regulation (Box 1). The domain architecture of class IB
ma membrane, but they play a crucial role in controlling regulatory subunits, by contrast, is not yet defined. The
intracellular signaling events. One of the most versatile catalytic subunits of the class I PI3K isoforms have dis-
signaling phosphoinositides is the lipid second-messenger tinctly different expression profiles, and p110a and p110b
phosphatidylinositol 3,4,5-trisphosphate (PIP3) produced are ubiquitously expressed whereas p110d and p110g are
from phosphatidylinositol 4,5-bisphosphate (PIP2) by the expressed primarily in hematopoietic cells [2].
class I PI3Ks. Class I PI3Ks are activated downstream of Class I PI3Ks play vital roles in regulating cellular
several signaling inputs, including G protein-coupled processes and are frequently misregulated in human dis-
receptors (GPCRs), receptor tyrosine kinases (RTKs), tyro- eases. The most prominent example is afforded by activat-
sine-phosphorylated adaptor proteins, and members of the ing somatic mutations of the gene for p110a (PIK3CA) that
Ras superfamily of small G proteins (Figure 1A). PIP3 are frequently found in human cancers [3,4]. Somatic
produced by PI3Ks is able to coordinate the recruitment mutations of other class I isoforms are not common, but
of effector proteins with PIP3-binding domains, including overexpression of any of these isoforms can induce onco-
several important signaling enzymes such as Ras super- genic transformation [5], and overexpression of these iso-
family guanine nucleotide exchange factors (GEFs), forms seems to play a crucial role in tumor growth in
GTPase-activating proteins (GAP), adaptor proteins, and particular tissues [2,6]. Intriguingly, there also appears
protein kinases. One of the best-studied PIP3 effector to be a role for PI3Ks in promoting tumorigenesis by
proteins is the Ser/Thr protein kinase Akt (also known affecting the tumor microenvironment [7] through a varie-
as protein kinase B, PKB). Recruitment of Akt to the ty of mechanisms including recruitment of immune cells
membrane through binding of its PH domain to PIP3 leads [8], preventing the action of cytotoxic CD8+ T cells [9], and
to its phosphorylation and activation by phosphoinositide- mediating angiogenesis [10]. Therefore, inhibitors against
dependent kinase (PDK1) and mammalian target of rapa- PI3Ks are currently in clinical trials for treating a variety
mycin complex 2 (mTORC2) [1]. These downstream effec- of cancers. PI3K inhibitors may play important roles in the
tors control a tremendous number of cellular responses treatment of other diseases in addition to cancer. For
Corresponding author: Burke, J.E. ([email protected]).
example, recent work has highlighted the discovery of
Keywords: phosphoinositide 3 kinases; lipid signaling; cancer; phosphoinositides; patients with primary immunodeficiencies who harbor
primary immunodeficiencies; lipid kinases. activating hereditary mutations in the gene encoding
0968-0004/ the p110d catalytic subunit (PIK3CD) [11,12], as well as
Crown Copyright ß 2014 Published by Elsevier Ltd. All rights reserved. http:// mutations in PIK3CA that is involved in mosaic over-
dx.doi.org/10.1016/j.tibs.2014.12.003
growth syndromes [13–15].

88 Trends in Biochemical Sciences, February 2015, Vol. 40, No. 2

 Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2
[(Figure_1)TD$IG]

(A) p110α p110δ RTKs p110β p110γ

BCR/TCR/TLRs FcγR
CD19/CD28 GPVIR
RTKs RTKs Integrin αIIbβ3 GPCRs RTKs, FcεR

pY Plasma membrane pY pY pY

PLCβ
GRB2 Ras Gβγ Ras
adaptor

P P Gβγ Gβγ Gα
P P P P PTEN P P P RASGRP4 P P
P SOS GTP ? GTP
P P P P
P P P P
P P P Dock180/Elmo1
P P p84/
p85 p110β Rac1 p110γ
p85 p110α p110δ p85 PIP2 PIP3 p101
GTP
PKCβ downstream of FcεR

(B) PDK1 PIP3 GAPs (Arf, Rho, Rac)

Cell molity
Cytoskeletal rearrangement
GEFs (Rac, Arf)

mTORC2 AKT GSK3

S6K mTORC1 Metabolism, cell cycle,
FOXO
growth, survival
4EBP1 MDM2
Protein synthesis TSC1
RHEB
TSC2
Ti BS

Figure 1. Signaling inputs into class I PI3K signaling and signaling outputs downstream of PIP3 production. (A) Class I PI3Ks are able to integrate a variety of direct and
indirect stimuli in their activation. Class I PI3Ks catalyze the formation of PIP3, and this is antagonized by the lipid phosphatase PTEN. PI3Ks can be activated downstream of
a variety of phosphorylated receptors (pY), GPCRs, and the Ras superfamily of GTPases. Phosphorylated receptors shown in bold are able to directly activate PI3K through
interactions with p85-like regulatory subunits. Receptors not in bold either activate indirectly or by an unknown mechanism. (B) PIP3 produced by PI3Ks recruits several
important signaling proteins, including Ras superfamily GEFs and GAPs, as well as the protein kinases PDK1 and PKB (also known as Akt). Phosphorylation of kinases and
transcription factors downstream of these enzymes play key roles in a variety of cellular signaling events. Lines shown in red in (A,B) represent phosphorylation events.
Abbreviations: BCR, B cell receptor; 4EBP1, eukaryotic translation initiation factor 4E-binding protein 1; FceR, Fc e receptor; FcgR, Fc g receptor; FOXO3, Forkhead box O3;
GAP, GTPase activating protein; GEF, guanine nucleotide exchange factor; GPCR, G protein-coupled receptor; GPVIR, ITAM-bearing collagen receptor glycoprotein VI;
GRB2, growth factor receptor-bound protein 2; GSK3, glycogen synthase kinase 3; MDM2, E3 ubiquitin-protein ligase; mTORC1 and 2, mammalian target of rapamycin
complex 1/2; PDK1, phosphoinositide-dependent kinase 1; PI3K, phosphoinositide 3 kinase; PIP3, phosphatidylinositol 3,4,5-trisphosphate; PKB, protein kinase B; PKCb,
protein kinase Cb; PLCb, phospholipase Cb; RasGRP4, RAS guanyl releasing protein 4; RTK, receptor tyrosine kinase; SOS, Son of Sevenless; S6K, P70-S6 kinase 1; TCR, T
cell receptor; TLR, Toll-like receptor; TSC1 and 2; tuberous sclerosis complex 1/2.

Because of the essential role of individual PI3K isoforms recent years have seen a steady advance in our under-
in signaling, in particular the crucial roles of p110a standing of the regulation of each class I PI3K isoform
and p110b in signaling downstream of insulin receptors downstream of activating stimuli, and proper integration
[16–18], broad-specificity PI3K inhibitors can have pro- of the isoform-specific signaling roles will be crucial in
found negative metabolic side effects [19]. To circumvent designing novel therapeutic strategies for targeting
these potential problems, considerable effort has been PI3K signaling in disease.
dedicated to developing isoform-specific PI3K inhibitors.
Initial clinical trial results using PI3K inhibitors have Isoform-specific signaling inputs into class I PI3Ks
shown limited single-agent activity [20,21], the exception Multiple signaling pathways can activate PI3Ks, including
being selective inhibitors of p110d in blood cancers. Idela- tyrosine phosphorylated receptors and their adaptors,
lisib (formerly known as CAL-101 or GS-1101), an inhibitor GPCRs, and the Ras superfamily (Figure 1). Each PI3K
of p110d, is one of the most selective PI3K inhibitors. It has isoform has a different ability to integrate signals from
recently been FDA approved for three blood cancers, in- these inputs, and many of the isoform-specific roles of
cluding chronic lymphocytic leukemia (CLL), and is in PI3Ks are dependent on their ability to successfully syner-
clinical trials for other blood cancers with preliminary gize inputs from multiple pathways. Detailed structural
results showing efficacy of these compounds against these and biochemical studies have defined the location and
cancer types [22–27]. One of the challenges of using PI3K mechanism of activation of many of these regulatory inputs
inhibitors in cancer is that frequently patients show a (Figure 2). Intriguingly, activating stimuli have been found
partial initial response, followed by tumor progression. to function by both direct and indirect activation mecha-
This is highlighted in a recent study where a patient with nisms, and this has revealed an unexpected level of com-
breast cancer containing a mutated p110a showed a clini- plexity in regulating PI3K function. A key example of this
cal response to a p110a-specific inhibitor; however, the is the activation of p110b downstream of Gbg subunits
cancer progressed through the convergent evolution of derived from GPCRs. Gbg subunits can directly activate
PTEN (phosphatase and tensin homolog) loss in different PI3K through interaction with p110b [29]; however,
metastatic sites [28]. With this intense clinical interest, Gbg subunits can also indirectly activate p110b through
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Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2

Box 1. Structural basis of PI3K regulation by regulatory subunits

The class IA catalytic subunits (p110a, p110b, p110d) consist of an All of the class IA catalytic isoforms are inhibited by the presence of
adaptor-binding domain (ABD), a Ras-binding domain (RBD), a C2 the iSH2 and the nSH2 domains of p85 [102,104,105]. This is mediated
domain, a helical domain, and a kinase domain that is composed of by contacts between the iSH2 and C2 [108], and contacts of the nSH2
N- and C-lobes similar to protein kinases (Figure I) [101]. All class IA with the C2, helical, and kinase domains [67,106]. The inhibitory
catalytic isoforms can associate equally with all of the class IA contact between the iSH2 and C2 domain is partially disrupted in
regulatory isoforms (p85a, p85b, p55a, p50a, p55g). The class IA p110b compared to both p110a and p110d [107]. Both p110b and
regulatory subunits contain two Src homology 2 domains, nSH2 and p110d make an extra inhibitory contact with p85 mediated by an
cSH2, separated by a coiled-coil domain known as the inter-SH2 interaction between the cSH2 of p85 and the C-terminal end of the
(iSH2). The iSH2 interacts tightly with the ABD of the catalytic subunit. kinase domain [103–105]. Phosphorylated tyrosines present on
The p85a and p85b subunits also contain an SH3 domain at their N- receptors and their adaptors activate the catalytic subunits by binding
terminus, along with a breakpoint cluster region homology domain to the SH2 domains of the regulatory subunit and disrupting the
(BH) flanked by two proline-rich regions. inhibitory interactions with the catalytic subunit.
The class IA regulatory subunits play three key roles: stabilizing the The class IB (p110g) catalytic subunit shares the same domains as
p110 subunit, inhibiting p110 basal lipid kinase activity, and facilitat- the class IA, except for the absence of an ABD at the N terminus. It
ing activation downstream of phosphorylated tyrosine motifs with can associate with either a p84 (also referred to as p87) or a p101
their nSH2 and cSH2 domains [102]. Structural data from X-ray regulatory subunit. They play important non-redundant roles in
crystallography and hydrogen–deuterium exchange mass spectro- coupling PI3K to upstream signaling pathways, including Ras and
metry (HDX-MS) along with biochemical studies have revealed that GPCRs [39], with p84 participating in a constitutively expressed PI3K
the different class IA isoforms have unique regulatory interactions complex, and p101 being part of an inducibly expressed PI3K
with p85 regulatory subunits [68,103–107]. complex [109].
[(Figure_I)TD$G]

(A) Class IA PI3Ks (C) Class IB PI3Ks

Kinase domain
p110α ABD RBD C2 Helical N-Lobe C-Lobe

Kinase domain
1 2 p110γ RBD C2 Helical N-Lobe C-Lobe
p85α,β SH3 P BH P nSH2 i cSH2

p50α p55α p55γ P nSH2 i cSH2

?
Kinase domain p84
p110β,δ ABD RBD C2 Helical N-Lobe C-Lobe p101

Key:
1 2 3
Binding interface
p85α,β SH3 P BH P nSH2 i cSH2 Inhibitory interface
p50α p55α p55γ P nSH2 i cSH2

(B)
Helical RBD Helical
p110 RBD

nSH2

120° ABD

nSH2

cSH2
Kinase
C2 iSH2 iSH2 p85 C2
Ti BS

Figure I. Domain architecture of class I phosphoinositide 3 kinases (PI3Ks) and the structural basis of their regulation by regulatory subunits. (A) The domain
architecture of all class IA PI3Ks and their p85-like regulatory subunits are shown, with both binding and regulatory interfaces shown according to the legend. (B)
Structural model of SH2-mediated inhibition of class IA catalytic subunits. This shows a model of p85 regulation of p110 catalytic activity composed of the structures of
the p110b catalytic subunit bound to the iSH2 and cSH2 domains from p85b [105], as well as the p110a catalytic subunit bound to the iSH2 and nSH2 domains [106]. (C)
The domain architecture of class IB PI3Ks as well as their proposed binding sites with p84 and p101 regulatory subunits.

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 Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2
[(Figure_2)TD$IG]

Ras
(p110α,δ) PKCβ
Ras Gβγ PKA
Rac/Cdc42 Gβγ
(p110β) (p110β)

S582 T1024
RBD C2 Helical N-Lobe C-Lobe p110γ
p110α,β,δ ABD RBD C2 Helical N-Lobe C-Lobe

S83 S361 S652,Y688, S690 p84

p85 (p85α) (p85α) (p85α)
SH3 P BH P nSH2 i cSH2 p101
regulatory 770–830
subunits ?

FXBL1 PKD Lck/Abl IKK Gβγ

14-3-3ζ PKC? pY/RTKs BRD7 NS1
(p85α) (free p85α) (p85β) (mediates (p85α) (p85α) (p85α)
Dependent on degradaon of
PKA phosphorylaon free p85β)

Key: Phosphorylaon site

Inhibitory interacon Inhibion mediated by phosphorylaon
Smulatory interacon Acvaon mediated by phosphorylaon
Ti BS

Figure 2. Class I PI3K (phosphoinositide 3 kinase) binding partners that regulate their activation. Schematic representation of identified class I PI3K binding partners is
shown, with their area of binding indicated by the arrow. Proteins represented in green are able to stimulate PI3K signaling, whereas those in red have inhibitory effects.
The arrows from the protein binding partners to PI3K represent the mechanism of activation or inactivation. Regulation through binding is colored in dark green for
activation, and red for inactivation. Regulation mediated through phosphorylation-dependent changes (phosphorylation site represented as red circle) are shown in purple
for inhibition, and light green for activation. A broken line indicates putative binding partners with an undetermined site of interaction. Both p84 and p101 are shown as grey
boxes because no domain annotation is available for these proteins. Abbreviations: see Figure 1 legend.

activation of Rac GTPases and subsequent activation of Sevenless and mediate p110a activation through Ras
p110b [30]. [31]. In insulin-responsive tissues, endothelial cells, epi-
Many activating stimuli are able to target multiple thelial cancer cells, and embryonic fibroblasts, the p110a
PI3K isoforms, and all class IA PI3Ks have the potential isoform primarily mediates signaling downstream of
to be activated downstream of phosphorylated receptors growth factor receptors and their adaptors [18,32,33]. In
and their adaptors through disrupting the SH2-mediated human breast-derived cells p110a is significantly less
inhibition of p85 regulatory subunits. Similarly, it might be abundant than p110b, but p110a is still required for
assumed that all class I PI3K isoforms could be equally signaling downstream of the epidermal growth factor re-
activated by Ras superfamily members through interac- ceptor (EGFR) [34]. The p110a isoform can also be activat-
tions with the Ras-binding domain (RBD). However, class I ed by a broad spectrum of Ras family members
isoforms show a high degree of sequence divergence in the [35,36]. Mice bearing germline mutations in the p110a
RBD, and different members of the Ras superfamily acti- RBD (T208D, K227A) that prevent it from binding to
vate different PI3K isoforms [30]. Crucial to understanding Ras were resistant to lung and skin cancer induced by
these isoform-specific activation mechanisms has been the mutant oncogenic Ras [35], indicating a crucial role of
generation of point mutations that have normal kinase p110a in mediating oncogenic transformation downstream
activity, but are unable to be activated downstream of of mutant Ras. Not only does this interaction play a role in
specific stimuli. These results are summarized below for tumorigenesis but it also plays a key role in tumor-induced
the signaling inputs into each of the class I PI3Ks. angiogenesis [37] and tumor maintenance because experi-
ments in an inducible mouse model revealed that blocking
Signaling inputs into p110a the ability of p110a to bind to Ras after tumor formation
The p110a isoform is activated by phosphorylated tyrosine led to tumor stasis and partial regression [38].
receptors, their adaptors, and Ras. Phosphorylated recep-
tors are able to activate p110a both directly and indirectly. Signaling inputs into p110b
They directly activate p110a through binding the p85 The p110b isoform is unique in that it is the only isoform
regulatory subunit, but indirect activation occurs through able to be activated downstream of both RTKs and
multiple mechanisms. Specifically, they can indirectly ac- GPCRs [39,40]. Most cellular experiments indicate that
tivate through either recruiting adaptor proteins, such as this isoform is less responsive to RTK signaling [40];
insulin receptor substrate 1 (IRS1) or GRB2-associated however, it is able to propagate signals downstream of
binder (GAB), which can directly recruit p85, or they can RTKs in cells deficient in other class IA isoforms [41].
recruit GRB2, which can activate the Ras GEF Son of This isoform is preferentially activated downstream of
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Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2

tyrosine phosphorylation of the Fcg receptor in neutro- Intriguingly, it has also been shown that p110d is crucial in
phils [42]. It also plays a key role in thrombosis in mediating signals downstream of GPCRs in natural killer
platelets through activation downstream of the integrin cells [49]; however, this is not through a direct interaction
aIIbb3 receptor [43], as well as the immunoreceptor and the exact mechanism is still unknown.
tyrosine-based activation motif (ITAM)-bearing glycopro-
tein VI receptor (GPVIR) [44,45]. The requirement of Control of PI3K signaling through inputs into p85
PI3Kb for activation downstream of these specific phos- The p85 regulatory subunits play key roles in regulating
phorylated tyrosine receptors and their adaptors has PI3K activity, and the structural and biochemical basis of
been suggested to be mediated through synergistic acti- this regulation is well established (Box 1). PI3K activity
vation with GPCRs. This isoform is activated by Gbg can be upregulated by p85 ligands that disrupt the in-
heterodimers resulting from GPCR activation, and Gbg hibitory interaction of p85 with p110, including the in-
interaction with p110b is mediated by the linker region teraction of p85b with the viral protein NS1, which is
between the C2 domain and helical domain (Figure 2) proposed to disrupt the nSH2 helical inhibitory interac-
[29]. Substitutions in this region (KK532–533DD) that tion [50].
prevented binding and activation by Gbg subunits pre- The p85 regulatory subunits of class IA PI3Ks have
vented oncogenic transformation driven by p110b. been proposed to exist both free or in complex with the
The p110b isoform is also unique among class IA PI3Ks p110 catalytic subunits. The role of free p85 regulatory
in that it is not activated downstream of the Ras subfamily. subunits in signaling has remained controversial: early
Instead, it is activated downstream of the Rho subfamily of p85 knockout models suggested that free p85 could com-
small G proteins, specifically by Cdc42 and Rac1 [30]. The pete with p110/p85 complexes for recruitment to RTKs
BH domain of free p85 has been previously suggested to [51]; however quantitative mass spectrometry experi-
interact with Cdc42; however, it has unambiguously been ments suggested that p110/p85 existed as an obligate
shown that the interaction between p110b/p85 and Cdc42 heterodimer [52]. It has been proposed that a small pool
or Rac1 is independent of the BH domain of p85, and is of free p85 modulates signaling through several mecha-
instead mediated by the RBD. Substitutions in the RBD of nisms, including activation of the lipid phosphatase PTEN
p110b (S205D/K224A) that decrease binding of Cdc42 and [53] and its BH domain that acts as a GAP for the small
Rac1 showed reduced signaling downstream of GPCRs. GTPase Rab5 [54,55].
The proposed mechanism for this Rac1–GPCR link is direct A possible explanation of these contradictory findings is
binding of the RacGEF Dock180 and its adaptor Elmo1 to that the free pool of p85 subunits is maintained at very low
Gbg subunits. Therefore, Gbg subunits from GPCRs are levels, and is maintained under tight control. The bromo-
not only able to directly activate p110b, but they also domain-containing protein 7 (BRD7) is able to bind to free
control the RacGEF upstream of small G proteins and p85a and sequester it to the nucleus, which leads to down-
therefore mediate p110b activation through both direct regulation of PI3K signaling through destabilization of
and indirect mechanisms. p110 catalytic subunits [56]. Another mechanism for tight
The intriguing observation that mutation of either the control of the concentration of free p85 subunits has also
Gbg interface or the Rac1/Cdc42 interface is able to abro- been described that involves the ubiquitin ligase-associat-
gate GPCR signaling through p110b may indicate that the ed F-box protein (FBXL-2) associating with free p85a and
activation of p110b requires coincident detection of multi- p85b [57]. FBXL-2 mediates the ubiquitylation and subse-
ple signaling inputs. The selective disruption of any of quent proteasomal degradation of free p85b when its Y655
these signaling inputs would then significantly decrease residue has been dephosphorylated by the protein phos-
signal output. However, this hypothesis will require test- phatase PTPL1. This type of mechanism may explain why
ing in the same experimental system. almost no free p85 subunits have been found in a variety of
cell types, and how signaling through a pool of free p85
Signaling input into p110d subunits would crucially depend on tight control of their
The p110d isoform is primarily expressed in hematopoi- synthesis and degradation. It is important to note that this
etic cells, and studies of kinase-dead knock-in PIK3CD control would not explain a mechanism where free p85
mice [46] have revealed key roles of this enzyme in could compete with p110/p85 complexes for recruitment to
mediating the immune responses of B and T cells. It RTKs, owing to the extremely low levels of free p85 com-
plays important roles in transmitting signals down- pared to p85 in complex with p110. However, the presence
stream of tyrosine phosphorylation of receptors and their of specific regulatory mechanisms that maintain free p85
adaptors in the immune system, including cytokine levels in cells suggest that free p85 may have important
receptors, Toll-like receptors, and the FceR [47]. The signaling roles.
p110d isoform can also be activated indirectly down- The activity of class IA PI3Ks can also be controlled by
stream of the B cell receptor (BCR) through the action post-translational modifications of the p85 regulatory sub-
of B cell adaptor for PI3K (BCAP), and by the T cell unit when in complex with p110 subunits (Figure 2). Protein
receptor through an unknown mechanism [47]. kinase C (PKC) stimulates phosphorylation of the nSH2 and
The p110d isoform is also activated downstream of a cSH2 domains at residues S361 and S652, and the IkB
subset of the Ras subfamily, specifically R-Ras and TC21 kinase (IKK) phosphorylates the cSH2 (S690). Phosphory-
(also known as R-Ras2) [36]. TC21 interacts with both T lation prevents these domains from binding to phosphory-
cell and B cell receptors and is necessary for the recruit- lated tyrosine receptors [58,59]. The cSH2 can also be
ment of p110d to antigen receptors in lymphocytes [48]. phosphorylated at Y688, and this was proposed to activate
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PI3K activity by facilitating an intramolecular interaction Isoform-specific signaling inputs into class I PI3Ks:
with nSH2 [60]. The discussion of phosphorylation sites summary
located in the SH2 domains as activating or inhibitory Extensive effort has gone into teasing out the isoform-
can be confusing because phosphorylation can not only specific signaling functions of each of the class I PI3Ks.
prevent inhibition by SH2 domains but can also prevent Intriguingly, this has revealed extensive crosstalk between
activation through decreased SH2-mediated recruitment to these signaling pathways, leading to an extraordinary
phosphorylated receptors. Phosphorylation of p85 outside of depth of complexity. This has revealed that many of the
the SH2 domains is also able to modify lipid kinase activity, isoform-specific functions of PI3Ks are crucially dependent
as phosphorylation of S83 in the SH3 domain of p85a by on the synergistic input of several upstream signals. This is
protein kinase A (PKA) mediates binding of 14-3-3z, and this not surprising because integrating a variety of upstream
association leads to increased membrane binding and PI3K inputs allows a much greater plasticity and responsiveness
activity [61]. of signaling. A crucial direction for future work is the
tissue-specific use of mutants deficient in specific signaling
Signaling input into p110g/p84 and p110g/p101 inputs, which will reveal the role of individual inputs into
The p110g isoform is activated downstream of both Ras each of the class I PI3K isoforms.
and GPCRs; however, recent studies have demonstrated
a complex interplay between these interconnected path- Misregulation of class I PI3Ks in disease
ways. Neutrophils from mice expressing RBD mutants of The important role of PI3Ks in cancer development was
p110g (T232D, K251A, K254S, K255A, K256A) that are highlighted by the discovery in 2004 that p110a is one of
unable to bind to Ras show decreased PIP3 levels and the most frequently mutated genes in cancer [3]. However,
decreased signaling downstream of GPCRs [62]. This several other diseases recently have been shown to be
interplay between GPCR and Ras signaling upstream mediated through mutation of PI3K isoforms, including
of p110g in neutrophils has been explained by the obser- primary immunodeficiencies and developmental disorders.
vation that Gbg heterodimers activate phospholipase C There have also been important developments in under-
isoforms (PLCb2/b3, which activate the specific RasGEF, standing the mechanism of how these mutations activate
RasGRP4, and therefore activate Ras [63]. An important PI3K signaling, as well as how PI3K can mediate cancer
and unappreciated role of signaling downstream of progression through modulating tumor microenviron-
the Ras superfamily is that different GEFs can be regu- ments and regulating immunological responses. Intrigu-
lated by many upstream signaling inputs, and these ingly, there are also indications that mutations within
same inputs may also be able to directly regulate PI3K PI3Ks can rewire signaling through modification of other
signaling. signaling pathways. The involvement of each PI3K isoform
One of the key mechanisms of regulating p110g and its in disease is summarized below with a particular focus on
response to activating inputs is through the presence of the role of PI3Ks in cancer and immune diseases.
different regulatory subunits. The heterodimer of p110g
with p84 shows decreased sensitivity to activation down- Misregulation of p110a
stream of Gbg subunits from GPCRs and requires Ras as Numerous activating mutations are distributed within
an indispensable activating partner [39]. The heterodimer each domain of p110a: two major hotspots are focused at
of p110g with p101 shows enhanced sensitivity to activa- the helical domain (E545K) and kinase domain (H1047R)
tion by Gbg subunits and does not require Ras for activa- (Figure 3A). These hotspot mutations activate by separate
tion. The molecular determinants of activation of the mechanisms. E545K breaks the inhibitory interaction be-
heterodimer of p110g with p101 have been determined: tween the nSH2 and the helical domain [67,68], and
the C2-helical linker in p110g is involved in an interactions requires signaling downstream of oncogenic Ras for trans-
with Gbg subunits as well as two regions in the C-terminal formation. The H1047R mutant remains dependent on p85
half of p101 [64]. Mutations at either of these sites caused a but no longer requires Ras for oncogenic transformation
significant decrease in activation downstream of Gbg sub- [69]. This is consistent with in vitro results showing that
units both in vitro and in vivo. the H1047R mutant can be hyperactivated downstream of
The p110g isoform can also be activated downstream of phosphopeptides derived from RTKs through increased
RTKs and Toll-like/IL-1 receptors (TLR/IL1Rs) in myeloid membrane affinity [68,70]. Activation of PI3K signaling
cells [8], and the high-affinity IgE receptor (FceRI) [65] in through both of these mutations causes large-scale
Mast cells. Activation downstream of these inputs is indi- changes in phosphorylation of the kinome, and concurrent
rect, and activation of RTK and TLR receptors leads to Ras- activation of numerous downstream signaling pathways
dependent activation of p110g, and activation downstream [71].
of FceRI involving Ca2+-dependent activation of PKCb Along with the hotspot mutations there are numerous
leads to phosphorylation of p110g at S582. Phospho-mi- clusters of mutations outside the hotspots, many of which
metic substitutions of this residue caused increased lipid are frequently found in endometrial cancer [72]. These
kinase activity and decreased interaction with p84 clusters are located at the interface of the adaptor-binding
[65]. The p110g isoform can also synergize with inputs domain (ABD) with the kinase domain, the linker between
from cAMP signaling because it can be phosphorylated by ABD and RBD, the C2–iSH2 interface, and the membrane
PKA at S1024, which downregulates PI3K activity and is interface of the kinase domain (Figure 3C). Many of these
crucial for maintaining contractility in cardiomyocytes clusters are distant from both the active site and the
[66]. proposed membrane interface. During activation of PI3K
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 Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2
[(Figure_3)TD$IG]
(A) Somac missense mutaons in PIK3CA (p110α) (B) Oncogenic truncaon mutaons in PIK3R1
2000
E545K H1047R SH3 P BH P nSH2
nSH2 i cSH2
cSH2
1000
Number of somac mutaons idenfied

60 N345K
PTEN acvaon, stabilizaon
30
0
0

00
10

20

30

40

50

60

70

80

90
SH3 P BH E160*

10
ABD RBD C2 Helical N-Lobe C-Lobe
SH3 P BH P nSH2 R348*

SH3 P BH P nSH2 L370 fs

SH3 P BH P nSH2 nSH2 i cSH2
cSH2
10 Somac missense mutaons in PIK3R1 (p85α)

Translocaon to nucleus, acvaon of JNK/ERK signaling

5

0
0

0
10

20

30

40

50

60

70

(C) Key: p110α regions that undergo conformaonal changes upon Key: p110α regions involved in
tyrosine phosphopepde/membrane binding membrane binding
Key:
Helical
RBD Mutaonal frequency Helical
>10 mutaons
E545K
>50 mutaons
nSH2 nSH2
>500 mutaons

ABD–RBD
linker
ABD kinase
interface
120°
ABD

H1047R
C2 Acve C2
N345K
C-lobe site
C2–iSH2 iSH2 Kinase iSH2
nSH2-helical
interface Helical interface
Helical
ABD–RBD SH3
BH
nSH2 p110α linker p110α nSH2
BH 120°
ABD nSH2
SH3 (p110α) p85α
C2 C2
Kinase
cSH2
iSH2 iSH2
cSH2 iSH2–C2
p85α interface
Ti BS

Figure 3. Location and mechanism of somatic mutations in PIK3CA [p110a, phosphoinositide-3-kinase (PI3K) catalytic subunit, a polypeptide] and PIK3R1 (p85a, PI3K
regulatory subunit 1a) in cancer. (A) Somatic mutations identified in cancer are shown on the primary sequence of both PIK3CA and PIK3R1. The frequencies of mutations
are from the catalog of somatic mutations in cancer database (COSMIC; http://cancer.sanger.ac.uk/cancergenome/projects/cosmic/). The sites of interaction of PIK3CA with
PIK3R1 are indicated with the location of somatic oncogenic mutations. Colored regions represent areas with increased exposure upon p110a activation (orange) or that
showed interactions with membranes (blue). (B) Oncogenic truncation mutations identified in PIK3R1 and their proposed mechanism of activation. All these truncation
mutants are unable to interact with the p110 catalytic subunit owing to the lack of the iSH2 domain. (C) Oncogenic mutations correlate with regions that either undergo
conformational changes or mediate membrane binding. The frequency of mutations in PIK3CA are mapped onto the structure of p110a in complex with the nSH2 and iSH2
of p85a [106] according to the key. Regions that showed conformational changes upon p110a activation are colored orange, and regions that showed interaction with
membranes are colored blue; mutants not at these regions are colored black [68].

the enzyme undergoes large allosteric conformational oncogenic mutations indicate that mutation of a single
changes and, intriguingly, many of these mutations are regulatory site, for example the ABD–RBD linker region,
located at the sites of these changes (ABD–RBD linker, C2/ causes a weakening of the distant C2/iSH2 interface
iSH2 interface, nSH2/helical interface, ABD/kinase inter- [68]. This suggests that all these mutations activate the
face) [68]. Experiments examining the dynamics of these enzyme by mimicking or enhancing specific conformational
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Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2

changes that are required for catalytic turnover of PI3K mediated by the BH domain of p85, leading to increased
enzyme activity. ubiquitylation and degradation of PTEN [79]. p85 poly-
Mutations throughout p110a directly activate the lipid peptides with truncation mutations within the nSH2 do-
kinase activity of PI3K; however, there has also been a main (R348* and L370fs) directly bind to the GTPases
question as to how oncogenic mutations might change the Cdc42 and Rac1, translocate to the nucleus, and activate
association with protein binding partners. A subset of extracellular signal regulated kinase (ERK)/c-Jun N-ter-
oncogenic p110a mutants including E545K are activated minal kinase (JNK) signaling [82]. This adds an additional
by binding to the non-phosphorylated form of IRS1 level of complexity in examining PI3K oncogenic mutations
[73]. This association is independent of the p85 subunit because mutations in p85a are able to activate through a
and can be disrupted by a peptide derived from the E545K mechanism not involving PI3K signaling.
helical domain. Mutations in p110a therefore not only Interestingly, downregulation of PI3K activity can also
activate lipid kinase activity, but they can also function be involved in disease. A substitution in the phosphopep-
by rewiring the upstream signaling pathways. tide binding site of the cSH2 in p85a (R649W) plays a role
There is an increasing body of evidence showing that in the rare complex developmental disorder SHORT syn-
mutations in p110a do not work alone in tumor develop- drome [short stature (S), hyperextensibility of joints or
ment. For instance, mice expressing H1047R p110a in hernia (inguinal) or both (H), ocular depression (O), Rieger
ovaries developed hyperplasias, but not tumors; however, anomaly (R), and teething delay (T)] [83]. This substitution
this substitution in combination with loss of PTEN led to causes decreased insulin-sensitivity by impairing the in-
the rapid development of tumors [74]. Mouse models of teraction of p110/p85a with phosphorylated IRS-1. This
H1047R p110a in breast cancer (reviewed in [75]) also shows that there are potential major side effects to globally
appear to require the presence of secondary mutations, dampening PI3K activity, and highlights the need for
with mice expressing H1047R in mammary tissue devel- isoform-specific PI3K inhibitors as therapeutics. A note
oping tumors in conjunction with several spontaneous of caution regarding PI3K modulation is raised by studies
secondary mutations that occur during tumorigenesis on the role of PI3Ks in development: activating PIK3CA
[76]. In mammary tumors driven by the overexpression mutations play key roles in overgrowth syndromes (see
of human epidermal growth factor receptor 2 (HER2), section on p110a), and inactivating mutations of p85a play
p110a is required for tumor development [77], and the key roles in developmental disorders, showing that a prop-
presence of H1047R enhances metastasis and causes re- er balance of PI3K signaling is crucial to normal develop-
sistance to standard anti-HER2 therapies [78]. ment.
The major focus on p110a misregulation in disease has
been its roles in cancer; however, several studies have Misregulation of p110b
identified mutations in p110a that play a key role in The p110b isoform plays a crucial role in the development
sporadic overgrowth syndromes, including megalence- of PTEN-null cancers in prostate cancer cells [16]; howev-
phaly-capillary malformation (MCAP) and megalence- er, other PTEN-deficient cancers can also be dependent on
phaly-polymicrogyria-polydactyly-hydrocephalus (MPPH) p110a [84]. The main role of p110b in cancer development
[13–15]. These mutations map to the same location as appears to be through the production of a high basal level of
somatic activating mutations identified in cancer, with PIP3, [16] which is sensitive to loss of PTEN, explaining the
similar hotspots in the helical and kinase domains. To- key role p110b in PTEN-deficient tumors. A very small
gether with mutations in PIK3CA, several mutations occur population of somatic mutations in p110b have been iden-
in other proteins involved in PI3K signaling, including tified in HER2-positive breast cancers [85], although at a
PTEN, AKT3, and p85b. Intriguingly, this opens up the markedly lower frequency than in p110a. Characterization
possibility of using PI3K inhibitors as potential therapeu- of a somatic mutation in the helical domain (E633K) in
tic agents for these syndromes. p110b revealed that it upregulates lipid kinase activity
independently of both RTK-derived phosphopeptide and
Misregulation of p85 Gbg activation [86], suggesting that a conformational
The p85a regulatory subunit is also frequently somatically change is induced by this mutation, possibly similarly to
mutated in human cancers [79–81], and most of these the cancer-linked mutations in p110a. However, given the
mutations upregulate class IA PI3K activity. Many point very low frequency of p110b mutations in cancer, it
mutations cluster at the interfaces that the iSH2 and nSH2 appears that their primary involvement in cancer is not
domains make with the p110 catalytic subunit (Figure 3A), dependent on somatic mutations, as seen for p110a.
and these interfaces are crucial in regulating class I PI3K
activity. A small number of cancer-linked p85 truncation Misregulation of p110d
mutants have been identified that are unable to bind to Multiple mutations in both PIK3CD (p110d) and PIK3R1
p110 catalytic subunits, and their mechanism of activation (p85a) have been identified in patients with primary immu-
has been proposed to be independent of PI3K catalytic nodeficiency diseases (Figure 4). A whole-exome study of
activity (Figure 3B). For instance, a truncation mutant in patients with primary immunodeficiencies (PID), which
the N-terminal portion of the BH domain (E160*) of p85a predispose them to chronic infections, revealed that about
found in endometrial tumors caused a decrease in the level 10% of these individuals had an E1021K substitution in
of the lipid phosphatase PTEN, which antagonizes PI3K p110d kinase domain [12]. Within families of these PID
signaling. It was proposed that this truncation upregulates patients, all individuals with the mutation had the disease,
PI3K signaling by disrupting a p85–PTEN complex that is and this has become known as activated PI3Kd syndrome
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 Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2
[(Figure_4)TD$IG]

(A) Mutaons in PIK3CD (p110δ)

Kinase domain
E334K E525K E1021K

p110δ ABD RBD C2 Helical N-Lobe C-Lobe

1 2 3

p85 SH3 P BH P nSH2 i cSH2

Δ434–475
(B) Mutaons in PIK3R1 (p85α)

RBD
Helical E1021K

E525K

nSH2

cSH2

Kinase

C2 E334K Δ434–475
iSH2
Ti BS

Figure 4. Primary immunodeficiency-linked mutations in (A) PIK3CD (p110d) and (B) PIK3R1 (p85a). The location of primary immunodeficiency mutations are shown on the
primary sequence of PIK3CD and PIK3R1. The regulatory inputs of p85 into p110d are shown according to the style of Box 1. The location of these mutations is mapped onto
a structural model of p110d bound to the nSH2, iSH2, and cSH2 of p85a.

(APDS). The mutation leads to constitutively increased changes the splicing of the p85a regulatory subunit mRNA,
p110d activity and is characterized by decreased numbers leading to deletion of residues 434–475 in the iSH2 domain
of B and T cells. Although the association with p110d is clear, [87].
the molecular mechanism giving rise to the activation-in- The same E1021K substitution as well as two others in
duced T cell death is not clear. An independent report the kinase domain have also been associated with diffuse B
showed dominant germline p110d substitutions in the C2, cell lymphomas [88]. These substitutions are analogous to
helical, and kinase domains (N334K, E525K, and E1021K) the somatic substitutions in p110a that are commonly
giving rise to a similar spectrum of disease symptoms. associated with human tumors. It is not clear whether
Intriguingly, these three residues all map to the regulatory wider screening for this substitution will uncover a strong
interfaces with p110d of p85 within its nSH2, iSH2, and association with lymphomas in APDS patients. Earlier
cSH2 domains (Figure 4B). Patients demonstrated a defi- work on patients with acute myelocytic leukemia (AML)
ciency in naive T cells and elevated senescent effector T cells [89] showed upregulation of PI3K activity, as well as
[11]. This study noted hyperactivation of mTOR associated increased expression of p110d and sensitivity to p110d
with enhanced glucose uptake, and this was proposed to inhibition, but among a cohort of 35 AML patients no
result in an increased number of cytotoxic CD8+ senescent T mutation in PIK3CD was found.
cells. Treatment of a patient with rapamycin, an inhibitor of B cell receptor (BCR) signaling controls the selection,
mTOR (a key effector downstream of PIP3), partially nor- proliferation, and differentiation of antibody-secreting
malized the T cell populations. Intriguingly, these muta- cells. As such, this signaling is fundamental to B cell
tions are not limited to the catalytic subunit, because malignancies. BCR signaling also participates in the func-
patients with primary immunodeficiencies have also been tion of self-reactive B cells involved in autoimmune dis-
identified who harbor a mutation in PIK3R1. This mutation eases. The p110d isoform couples BCR activation to B cell
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Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2

survival and migration, and early clinical trials indicate There is increasing evidence that PI3Kg has important
that p110d inhibitors such as Idelalisib may be important functions in metabolic diseases. Mice without functional
in treating both lymphoid malignancies and autoimmune PI3Kg are leaner, with protection from insulin resistance
diseases [90]. and fatty liver [98]. It has been proposed that because
The success of p110d inhibitors in cancer treatments obesity is associated with chronic inflammation, and
may be attributable to their effects on the tumor microen- PI3Kg plays a prominent part in obesity-associated inflam-
vironment. There is robust crosstalk between chronic lym- mation and insulin resistance, PI3Kg might be a drug
phocytic leukemia (CLL) cells and associated stromal cells target for obesity. Support has been provided by a recent
in the microenvironment which support malignant B cell study which showed that mice with genetically or pharma-
growth and increase drug resistance. Disrupting this cross- cologically lowered p110g and p110b kinase activity had a
talk between CLL cells and their microenvironment repre- dramatic decrease in fat mass, and this was mediated
sents a new therapeutic strategy that has interesting through modifying the sympathetic drive in the brain [99].
potential [91,92].
The prominent role of p110d in hematopoietic cells has Misregulation of class I PI3Ks in disease: a summary
led to efforts targeting this isoform in hematological can- PI3Ks have been major targets for therapeutic interven-
cers; however, increased p110d expression also has been tion owing to their key roles in the development of a variety
observed in solid tumors such as glioblastoma, prostate of cancers. An important recent development has been the
carcinoma, neuroblastoma, and breast cancer (reviewed in discovery of the role that class I PI3Ks play in other human
[93]). A microRNA (miR-663) that specifically targets diseases including developmental disorders and primary
p110d is a potent tumor suppressor in glioblastomas, immunodeficiencies. There has also been extensive re-
and is positively correlated with patient survival [94]. This search into the role of PI3Ks in mediating immune
suggests a possible use of p110d-specific inhibitors in these responses, and into the key role they play in controlling
diseases; however, the exact mechanisms of how decreased the tumor microenvironment. Of particular interest is the
p110d expression mediates increased survival is unknown, possibility of targeting PI3K in cancer therapy both
and may possibly be through decreased PI3K signaling in through targeting PI3K signaling in cancer cells them-
the tumor or through changes in the tumor microenviron- selves, as well as by targeting PI3K signaling within the
ment. tumor microenvironment to mediate anti-tumorigenic im-
A particularly exciting recent development has been the mune responses. Many of the initial single-use clinical
discovery that p110d plays a key role in suppressing the trials of PI3K agents have given an underwhelming thera-
cytotoxic ability of CD8+ regulatory T cells [9]. The sup- peutic response; however, given the plethora of new knowl-
pression of p110d activity is able to activate the antitumor edge on the mechanism of PI3K signaling, there are many
potential of these CD8+ cells, and thus can induce tumor exciting new possibilities for targeting PI3K in cancer and
regression. The p110d isoform also plays a role in mediat- other diseases. Key to this strategy will be designing novel
ing the activity of these CD8+ cells, and a potentially combinations of PI3K inhibitors with other signaling tar-
interesting possibility is therefore to specifically downre- gets, as well as tuning PI3K inhibitors that are able to
gulate the activity of p110d only in regulatory T cells, manipulate the immune response to tumors.
which would turn off the immunosuppressive function of Many oncogenic mutations in PI3K have been identi-
p110d while maintaining the full activity of CD8+ cells. fied, and an intriguing discovery has been how oncogenic
mutations are able to rewire signaling pathways. The
Misregulation of p110g unexpected discoveries that p85 truncation mutants acti-
PI3Kg is a long-established target for anti-inflammatory vate JNK/ERK signaling, and that the E545K substitution
and autoimmune therapies, and a newly described PI3Kg- in the helical domain of p110a rewires activation down-
specific inhibitor has been instrumental in discovering a stream of IRS1, underscore the importance of defining the
role for p110g in TH17 cell differentiation [95]. However, molecular basis of activating PI3K mutations.
more recent work has brought to light its roles in cancers Another major problem with PI3K pathway inhibitors is
and in metabolic diseases. the development of drug resistance after initial response.
Both the p110g catalytic subunit and the p101 regula- An intriguing possibility for individualized therapy in this
tory subunit play roles in tumor development. Depletion of situation is to culture circulating tumor cells (CTCs) from
p110g or p101 inhibits metastasis and primary tumor individual patients and use them to test the drug suscep-
formation in mouse epithelial cell cancers [96]. The gene tibility of the tumors throughout the treatment regimen to
for the p101 regulatory subunit is a common insertion site identify the best combination of therapeutic agents [100].
for murine leukemia virus [97], and this led to the obser-
vation that p101 overexpression in T cells increases PI3Kg Concluding remarks
signaling and protection from apoptosis. Similarly to Significant progress has been made in deciphering the
p110d, p110g also seems to play key roles in modulating exact inputs into different class I PI3K isoforms, as well
the tumor environment, and p110g mediates myeloid cell as their involvement in a variety of human diseases.
recruitment to tumors, leading to tumor inflammation and However, this has revealed unexpected complexities in
cancer progression [8]. Pharmacological or genetic block- the regulation and signaling downstream of class I PI3Ks.
ade of p110g in this mice model suppressed inflammation, Many unanswered questions remain (Box 2), and answer-
growth, and metastasis of implanted and spontaneous ing these will be essential to fully realizing the therapeutic
tumors. potential of targeting PI3K signaling in disease. The
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Feature Review Trends in Biochemical Sciences February 2015, Vol. 40, No. 2

Box 2. Outstanding questions 14 Rivière, J-B. et al. (2012) De novo germline and postzygotic mutations
in AKT3, PIK3R2 and PIK3CA cause a spectrum of related
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 What is the importance of feedback in PI3K signaling? PIK3CA cause CLOVES syndrome. Am. J. Hum. Genet. 90, 1108–
 What is the mechanism of activation of disease-linked mutations 1115
in each PI3K isoform? 16 Jia, S. et al. (2008) Essential roles of PI(3)K-p110beta in cell growth,
 How can PI3K isoform-selective inhibitors be correctly applied in metabolism and tumorigenesis. Nature 454, 776–779
cancer therapy in combination with other targeted therapeutics? 17 Ciraolo, E. et al. (2008) Phosphoinositide 3-kinase p110beta activity:
 How can modification of the tumor microenvironment through key role in metabolism and mammary gland cancer but not
inhibition of PI3K activity be harnessed for cancer therapy? development. Sci. Signal. 1, 1–13
18 Knight, Z. et al. (2006) A pharmacological map of the PI3-K family
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19 Smith, G.C. et al. (2013) Extended treatment with selective PI 3-
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20 Fruman, D.A. and Rommel, C. (2014) PI3K and cancer: lessons,
covery of the role of PIK3CD in mediating immune
challenges and opportunities. Nat. Rev. Drug Discov. 13, 140–156
responses against tumors will only enhance interest in 21 Rodon, J. et al. (2013) Development of PI3K inhibitors: lessons learned
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23 Gopal, A.K. et al. (2014) PI3Kd inhibition by idelalisib in patients with
specific PI3K isoforms that are deficient in individual sig- relapsed indolent lymphoma. N. Engl. J. Med. 370, 1008–1018
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the exact mechanism of how PI3Ks are involved in disease. lymphocytic leukemia. N. Engl. J. Med. 370, 997–1007
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yond inhibitors of the conserved ATP-binding pocket. 26 Brown, J.R. et al. (2014) Idelalisib, an inhibitor of phosphatidylinositol
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Acknowledgments Blood 123, 3390–3397
J.E.B. is supported by a discovery research grant from the Natural 27 Flinn, I.W. et al. (2014) Idelalisib, a selective inhibitor of
Sciences and Engineering Research Council of Canada (NSERC-2014- phosphatidylinositol 3-kinase-d, as therapy for previously treated
05218). R.L.W. is funded by the British Heart Foundation (PG11/109/ indolent non-Hodgkin lymphoma. Blood 123, 3406–3413
29247) and the MRC (file reference U10518430). 28 Juric, D. et al. (2014) Convergent loss of PTEN leads to clinical
resistance to a PI(3)Ka inhibitor. Nature Published online November
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