modulation of t cell response by targeting the pi3k akt...

Modulation of T cell response by targeting the PI3K‐Akt pathway

John Janik, M.D.Samir Khleif LaboratoryAugusta University

PI3KδPI3Kδ PI3KβPI3KβPI3KαPI3Kα PI3K

PI3K-Akt Signaling Pathway in T cells

Conventional T cells

Regulatory T cells

?

Normal Cell

T‐Cell

Tregs

T‐Cell

Tregs

CD4+Foxp3‐

CD4+Foxp3+

PI3K

PIP2 PIP3

PTEN, SHIP‐1 and ‐2

Akt

PIP3

PDK‐1PP

T308S473

mTOR

S6K1/2

P

S6

Proliferation

TCR

Stimulation

Effects of PI3K‐Akt pathway inhibition in Tregs vs. Tconv cells

PI3K

PIP2 PIP3


WM

Akt

PIP3

PDK‐1PP

T308S473

mTOR

S6K1/2

P

S6

TCN

Proliferation

TCR

Stimulation


PI3K

PIP2 PIP3


WM

AktPIP3

PDK‐1PP

T308S473

mTOR


TCR Stimulation ‐ + + +Treatment None None DMSO WM

Tconv

Treg

TCR

StimulationpAkt 473

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

US NT DMSO WM

pAkt 473/A

ktCD25‐ (Tconv)CD25+ (Treg)

S6K1/2P

S6

S6 phosphorylation is diminished in mouse Treg in presence PI3K/AKT inhibitors

Abu Eid R.et al, CIR, 2014

Effects of PI3K‐Akt pathway inhibition on the TCR/IL2 Induced proliferation of Tregs vs. Tconv cells

PI3K

PIP2 PIP3


WM

AktPIP3

PDK‐1PP

T308S473

mTOR

S6K1/2P

S6

TCN

Proliferation

TCR

Stimulation

Inhibition of PI3K‐Akt pathway in vivo

PI3K/ Akt pathway is essential for the in vivo maintenance of Treg in naïve tumor free mice

Abu Eid R.et al, CIR, 2014 ‐7 ‐5 ‐3 0

WM/TCN/MK2206 Collect splenocytes

PI3K‐Akt pathway inhibition indirectly decrease tumor growth

CT26 tumor growth

0

100

200

300

400

500

600

700

3 7 10 13 17 20 24

Days post tumor inoculation

Tumor size (m

m2)

PBS

0

Tumor implantation


CT26 tumor growth

0

100

200

300

400

500

600

700

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 )

PBSPC61

‐4 0

Tumor implantationPC61



CT26 tumor growth

0

100

200

300

400

500

600

700

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 ) PBSPC61DMSO

‐4 0

Tumor implantationPC61



CT26 tumor growth

0

100

200

300

400

500

600

700

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 )PBSPC61DMSOWM

‐7 ‐5 ‐3 0

Tumor implantationWM



CT26 tumor growth

0

100

200

300

400

500

600

700

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 )PBSPC61DMSOWMTCN

‐7 ‐5 ‐3 0

Tumor implantationTCN



0 Tumor size

EL4

0

100

200

300

400

500

600

700

800

900

3 7 10 13 17 20 24Days post tumor implantation

Tum

or S

ize

(mm

2)NT-PBS

NT‐PBS


PI3K‐Akt pathway inhibition of tumor growth is Treg‐dependent

‐7 ‐5 ‐3 0 Tumor size

EL4

WM

0

100

200

300

400

500

600

700

800

900


Tum

or S

ize

(mm

2)NT-PBSWM-PBS

* p<0.01

NT‐PBS

WM‐PBS



‐4 0 Tumor size

PC61 EL4

0

100

200

300

400

500

600

700

800

900


Tum

or S

ize

(mm

2)NT-PBSPC61-PBSWM-PBS

* p<0.01

NT‐PBSPC61‐PBSWM‐PBS



‐4 0 4 Tumor size

PC61 EL4 Treg

0

100

200

300

400

500

600

700

800

900


Tum

or S

ize

(mm

2)NT-PBSPC61-PBSWM-PBSPC61-Treg

* p<0.01

NT‐PBSPC61‐PBSWM‐PBSPC61‐Treg



‐7 ‐5 ‐3 0 4 Tumor size

EL4 Treg

WM

0

100

200

300

400

500

600

700

800

900


Tum

or S

ize

(mm

2)

NT-PBSPC61-PBSWM-PBSPC61-TregWM-Treg

* p<0.01

NT‐PBSPC61‐PBSWM‐PBSPC61‐TregWM‐Treg

Any effects on vaccine efficacy in vivo?


0

50

100

150

200

250

300

350

UT DMSO WM TCN

Spots p

er m

illion

E7 re‐stimDMSO re‐stim

‐7 ‐5 ‐3 0 14

E7 Vx Collect splenocytes

No Vx E7 Vx

* *** P<0.05; ** P<0.01

WM/TCN

PI3K‐Akt inhibition enhances vaccine efficacy


TC1 tumor growth

0

100

200

300

400500

600

700

800

900

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 )

DMSO-DMSO

Inhibition of PI3K‐Akt synergistically enhances anti‐tumor immune responses


TC1 tumor growth

0

100

200

300

400500

600

700

800

900

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 )DMSO-E7DMSO-DMSO



TC1 tumor growth

0

100

200

300

400500

600

700

800

900

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 ) WM-DMSODMSO-E7DMSO-DMSO



TC1 tumor growth

0

100

200

300

400500

600

700

800

900

3 7 10 13 17 20 24


Tum

or s

ize

(mm

2 )WM-E7WM-DMSODMSO-E7DMSO-DMSO



Akt inhibition by MK‐2206 enhances the anti‐tumor therapeutic effect of tumor‐specific vaccine..


Akt inhibition by MK‐2206 enhances the anti‐tumor immune response of tumor‐specific vaccine


WM differentially affects proliferation of human Tregs and Tconv cells


A phase Ib /II trial of pembrolizumab and idelalisibin patients with non‐small cell lung cancer (NSCLC)

Metastatic NSCLC • Progressed on platinum doublets• OR, Not able to tolerate chemo or unwilling to undergo chemotherapy

• OR EGFR, ALK, ROS1 mutations with disease progression on small molecule inhibitors

• may have also failed prior PD‐1/PD‐L1 blockade

Keytruda 200 mg Q3WZydelig 150 mg BID PO

Keytruda 200 mg Q3WZydelig 150 mg BID PO

Phase II(A) 50 PD1N patients (B) 20 PD1P patients

=2/6 DLTone dose lower

For Phase II

lower level if>2/6 DLT

<=1/6 DLT

Phase Ib

A phase Ib /II trial of pembrolizumab and idelalisibin patients with non‐small cell lung cancer (NSCLC)

Clinical Trials• Vaccine in combination with AKT inhibitors• AKT inhibitor in combination with anti‐PD1

•Samir Khleif•Shamim Ahmad•Pandelakis Koni•Mikayel Mkrtichyan•Rajeev Shrimali•Rasha Abu-Eid•Abdeljabar El Andaloussi•Atbin Doroodchi•Zuzana Berong•Mason Webb•Abdulmoneim Algady

•Tanusree Sen•Stephanie Zdaniv•Michiko Shimoda•Raed Samara•Osama Rahma

• Collaborators:– Theresa Whiteside, U Pitt– Jay Berzofsky, NCI– Nadia Tarasova, NCI– Jeff Schlom, NCI– Micheal Schickler, CureTech– Ramy Ibrahim, AZ– Ross Stewart, AZ– Linda Liu, Medimmune– Sol Langerman Medimmune

Acknowledgments

modulation of t cell response by targeting the pi3k akt...

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