PI3K signalling in immune cells

Bart Vanhaesebroeck, Klaus Okkenhaug and Christian Rommel

Many classes of immune-cell receptors relay intracellular signals considered to be attractive pharmacological targets in inflamma-
through phosphoinositide 3-kinases (PI3Ks), which culminates in tory and allergic diseases, and transplantation. Other PI3K isoforms
a broad variety of cell biological effects. Of the eight catalytic iso- are also expressed by leukocytes, but their roles in immune signal-
forms of PI3K that exist in mammals, p110δ and p110γ are highly ling are largely unexplored. This Poster displays the four immune-
expressed in all leukocyte subtypes. Pharmacological and mouse cell types — T cells, B cells, mast cells and neutrophils — in which
gene-targeting studies have identified these PI3K isoforms as p110δ and p110γ signalling and their biological consequences are
key enzymes in immune signalling, and p110δ and p110γ are best characterized.

BCR RTK
CD19
GPCR
CD4 or CD8 TCR IL-4R Plasma membrane
T cell B cell
CD3 Igβ Igα BAFFR PtdIns(4,5)P2 PtdIns(3,4,5)P3 PtdIns(4,5)P2
TRIM
CD28 or ICOS ζ RAS RAS β γ αi

LAT
p85 p110δ p101/ p110γ
Y P PTEN, SHIP p84
ZAP70 PLCγ1 PLCγ2 BCAP IRS2
VAV
LCK Ca2+ SLP65 TLR4 Class IA PI3K Class IB PI3K

DS
SYK

GA
VAV BTK Receptors RTKs, cytokine receptors, TLRs, antigen receptors, GPCRs, such as CCRs, adenosine receptors,

MD
FYN SLP76 TLR3 or TLR9 LYN FcRs, integrins P2YR

2
IL-2R ITK CD14
PKCβ Catalytic PI3K subunits p110α, p110β, p110δ p110γ
Regulatory PI3K subunits p85α, p85β, p55α, p55γ, p50α p101, p84 (also known as p87)
p85 p110δ
88 Receptor-associated adaptor IRS2, GAB2 G-protein βγ
p85 p110δ D
My
proteins
PKCθ

88
yD
M
AKT p50 p65 p50 p65 AKT
Class I PI3Ks: linking cell-surface receptors to GTPases include the P-REX family of RAC-GEFs,
FOXO
the production of a lipid second messenger the cytohesin family of ARF-GEFs and the
OX40 NF-κB NF-κB Class IA PI3Ks are heterodimeric proteins that centaurin family of ARF-GAPs. ARAP3 has
bind, through their regulatory subunits (known recently been identified as a PtdIns(3,4,5)P3-
MTOR FOXO GSK3
as p85 subunits), to tyrosine-phosphorylated and RAP-dependent GAP for RHOA. The roles
RAPTOR MTOR recognition motifs in the cytoplasmic domains of these small GTPase regulators in the immune
of RTKs or receptor-associated adaptor proteins. system are largely unknown.
Growth, survival, Growth, survival, proliferation, RAPTOR Other pathways
CCR7 or After cell stimulation, class I PI3Ks are recruited to
CXCR4 proliferation, differentiation, chemotaxis, glucose metabolism, (such as DOCK2)
β trafficking, cytokine antibody production, suppression G-protein the plasma membrane, where they phosphorylate p110δ versus p110γ
p101 p110γ RAC PtdIns(4,5)P2 to generate the lipid second Studies involving the blockade of p110δ or
γ synthesis and glucose of RAG expression and
β messenger PtdIns(3,4,5)P3. p110γ function, either by gene targeting or by
metabolism class-switch recombination CXCR5
G-protein
RAC p85 p110δ γ The class IA isoforms of PI3K (p110α, p110β and pharmacological inhibition, have confirmed
Other pathways (such as DOCK2) p110δ) signal downstream of tyrosine kinases important roles for both these PI3Ks in regulating
Biological effects and RAS. The only class IB PI3K isoform, p110γ, the immune system and in immune responses
is recruited to cytoplasmic protein complexes of in vivo. However, it is not clear if immune cells can
PKCζ PDKI GPCRs by interacting directly with G-protein βγ sense whether PtdIns(3,4,5)P3 is produced by
FAK subunits. This binding is facilitated by the class IB p110δ or by p110γ, or whether PtdIns(3,4,5)P3
Phagocytosis, Migration,
IL-3R MTOR Survival, proliferation, adhesion, oxidative burst, cell polarization, p85 p110δ SRC PI3K regulatory subunit (p101 or p84 (also known produced by both PI3K isoforms has the same
migration, cytokine synthesis, cytokine synthesis, adhesion, Integrin as p87)). RAS can also activate p110γ. signalling output.
GSK3 FOXO RAPTOR AKT
degranulation and secretion chemotaxis and membrane and Although the topic is not addressed in this Some pathways, such as signalling downstream
GAB2 motility actin dynamics PTK
Poster, distinct functions mediated by the different of the antigen receptors in lymphocytes and mast
Small p85 isoforms (p85α and p85β) have been revealed cells, are highly sensitive to inhibition of p110δ
AKT Actin assembly GTPases by knocking out the genes encoding these but not p110γ. By contrast, chemokine-receptor
NF-κB
machinery regulatory subunits in mice. For example, p85α signalling in most cells (such as neutrophils,
p50 p65 is the main regulatory isoform for many B-cell monocytes and macrophages) is mainly regulated
KIT AKT GEFs/ responses, but its role is more limited or by p110γ rather than by p110δ, although B-cell
Actin redundant in T cells and mast cells. chemotaxis in response to CXCL13 is more
PTEN GAPs
p85 p110δ Ca2+/ dependent on p110δ than on p110γ.
PKC
PI3K effectors Although they often operate in apparently
SOD
Mast-cell p101 p110γ γ The activation of all class I PI3Ks leads to increased parallel pathways, p110δ and p110γ can also
granule PTP H2O2 O2• β levels of the lipid second messenger PtdIns(3,4,5)P3 operate in the same pathway, as exemplified by
Secondary
response p85 p110δ at the cell membrane, which serves as a docking fMLP-stimulated PI3K activity in human neutrophils
platform for pleckstrin-homology-domain- primed with TNF. Stimulation of TNF-primed
GAB2 Primary CCR containing proteins, which then become activated. neutrophils with fMLP results in the biphasic
LYN SYK GADS response Phox Phox This leads to a cascade of phosphorylation events and activation of PI3K, with the first phase being
VAV p101 p110γ p101 p110γ p67 p47 NADPH NADP+ protein–protein interactions between downstream mainly p110γ dependent and the second phase
FYN SLP76 RAC
γ β γ β Phox targets to control multiple biological processes. being mainly p110δ dependent. The second phase
PLCγ1 BTK PTK
NTAL

P-REX1 p40 The serine/threonine kinase AKT is only one of PI3K activation is triggered by the first phase.
Phox p22

Adenosine Phox Recent work has identified the p110δ and

gp91 O2• O2• of several sensors of PtdIns(3,4,5)P3 in a cell. In
leukocytes in particular, tyrosine kinases, such as p110γ isoforms of PI3K as promising targets for
LAT

Oxidative members of the BTK family, are other key targets immune-mediated disorders, and several drug-
α β γ burst of PtdIns(3,4,5)P3. Other effectors include adaptor discovery programmes have been initiated to
A3AR
fMLPR Phox complex proteins, such as GAB1 and GAB2, and the GEFs develop small molecule inhibitors of these
FcεRI TNFR1
and GAPs for small GTPases, such as RAC and the isoforms as potential therapeutics for a wide
Mast cell Degranulation Neutrophil ARF-family members. These regulators of small range of human diseases.

Merck Serono is a global biotechnology leader, with sales in over 90 countries. The Company is the A3AR, adenosine receptor 3A; ARAP3, ARF–GAP, RHO–GAP, ankyrin repeat and pleckstrin-homology-domains- MyD88, myeloid differentiation primary-response gene 88; NF-κB, nuclear factor-κB; NTAL, non-T-cell activation Further reading Hirsch, E. et al. Thromb. Haemost. 95, Vanhaesebroeck, B., Ali, K., Bilancio, A., Christian Rommel is at Merck Serono
Vanhaesebroeck, B. et al. Annu. Rev. 29–35 (2006). Geering, B. & Foukas, L. C. Trends Biochem. International S.A., 9 Chemin des Mines,
world leader in reproductive health, with Gonal-f®, Luveris® and Ovidrel®/Ovitrelle®. It has strong containing protein 3; ARF, ADP-ribosylation factor; BAFFR, B-cell-activating factor receptor; BCAP, B-cell linker; O2•, superoxide; P2YR, purinergic receptor P2Y; PDK1, 3-phosphoinositide-dependent protein kinase 1;
Biochem. 70, 535–602 (2001). Ruckle, T., Schwarz, M. K. & Rommel, C. Sci. 30, 194 –204 (2005). 1211 Geneva, Switzerland. e-mail:
market positions in neurology, with Rebif®, as well as in metabolism and growth, with Saizen®, PI3K adaptor; BCR, B-cell receptor; BTK, Bruton’s tyrosine kinase; CCR, CC-chemokine receptor; CXCL, Phox, phagocyte oxidase; PKC, protein kinase C; PLC, phospholipase C; P-REX, PtdIns(3,4,5)P3-dependent RAC Okkenhaug, K. & Vanhaesebroeck, B. Nature Rev. Drug Disc. 5, 903–918 [email protected]
Serostim® and Zorbtive™. The Company has recently entered the psoriasis area with Raptiva®. CXC-chemokine ligand; CXCR, CXC-chemokine receptor; DOCK2, dedicator of cytokinesis 2; FAK, focal exchanger; PtdIns(4,5)P2, phosphatidylinositol-4,5-bisphosphate; PtdIns(3,4,5)P3, phosphatidylinositol-3,4,5- Nature Rev. Immunol. 4, 317–330 (2003). (2006). Affiliations and acknowledgements
We greatly appreciate K. Ali, H. Ji, M. Camps,
Wymann, M. P. et al. Biochem. Soc. Trans. Hawkins, P. T. et al. Biochem. Soc. Trans. Bart Vanhaesebroeck is at the Ludwig
Merck Serono’s research programs are focused on growing these businesses and on establishing adhesion kinase; FcεRI, high-affinity Fc receptor for IgE; fMLPR, N-formyl-methionyl-leucyl-phenylalanine trisphosphate; PTEN, phosphatase and tensin homologue; PTK, protein tyrosine kinase; PTP, protein tyrosine T. Ruckle and M. K. Schwarz for their help
31, 275–280 (2003). 34, 647–662 (2006). Institute for Cancer Research, 91 Riding
receptor; FOXO, forkhead box O; GAB, GRB2-associated binding protein; GADS, GRB2-related adaptor phosphatase; R, receptor; RAG, recombination-activating gene; RAPTOR, regulatory associated protein of with the Poster, and all our collaborators for
new therapeutic areas, including oncology and autoimmune diseases. Deane, J. A. & Fruman, D. A. Annu. Rev. Okkenhaug, K., Ali, K. & House Street, London, W1W 7BS, UK.
their support.
protein; GAP, GTPase-activating protein; GEF, guanine-nucleotide-exchange factor; GPCR, G-protein- MTOR; RHOA, RAS homologue gene-family member A; RTK, receptor tyrosine kinase; SHIP, SRC-homology-2- Immunol. 22, 563–598 (2004). Vanhaesebroeck, B. Trends Immunol. e-mail: [email protected]
For more information, please visit: www.merckserono.net coupled receptor; GRB2, growth-factor-receptor-bound protein 2; GSK3, glycogen-synthase kinase 3; H2O2, domain-containing inositol-5-phosphatase; SLP, SRC-homology-2-domain-containing leukocyte protein; SOD, Wetzker, R. & Rommel, C. Curr. Pharm. 28, 80–87 (2007). Klaus Okkenhaug is at the Laboratory of Edited by Sharon Ahmad and Lucy Bird;
Design 10, 1915–1922 (2004). Rommel, C., Ji, H. & Camps, M. Lymphocyte Signalling and Development, designed by Neil Smith.
hydrogen peroxide; ICOS, inducible T-cell co-stimulator; IL, interleukin; ITK, IL-2-inducible T-cell kinase; superoxide dismutase; SYK, spleen tyrosine kinase; TCR, T-cell receptor; TLR, Toll-like receptor; TNFR1, tumour- Wymann, M. P. & Marone, R. Curr. Opin. Nature Rev. Immunol. 9 February 2007 Babraham Institute, Cambridge, CB2 4AT, UK. © 2007 Nature Publishing Group.
IRS2, insulin receptor substrate 2; LAT, linker for activation of T cells; MTOR, mammalian target of rapamycin; necrosis factor receptor 1; TRIM, TCR-interacting molecule; ZAP70, ζ-chain-associated protein kinase of 70 kDa. Cell Biol. 17, 141–149 (2005). (doi:10.1038/nri2036). e-mail: [email protected] www.nature.com/reviews/immunol