SARC Sarcoma SPORE

Raphael Pollock, MD, PhD

Principal Investigator

SARC Sarcoma SPORE Awarded September 2012 Multicenter collaboration

Ohio State University MD Anderson Cancer Center Dana Farber/Harvard University of Michigan Children’s National Hospital Columbia University University of Utah National Cancer Institute Cancer Research and Biostatistics (CRAB)

Project 1Histone deacetylase inhibitor (HDACi)-based therapeutic strategies for the treatment of genetically complex STS

Co-Leaders Raphael Pollock, MD, PhD (Ohio State) Shreyaskumar Patel, MD (UTMDACC)

Specific Aims Aim 1: To evaluate the activity of an HDACi/doxorubicin

combination in patients with metastatic LMS Aim 2: To examine the role of autophagy as a novel

process contributing to HDACi tolerance Aim 3: To determine the impact of HDAC8 blockade on

STS growth in vitro and in vivo

Specific aim 1 Phase 2 of

mocetinostat and gemcitabine in chemorefractory LMS

Study cohort > age 18 Metastatic LMS Progression after

treatment with a gemcitabine-based regimen

Sponsor• SARC

N=30

Specific aim 2 Screeened HDACi for

pan-HDAC isoform inhibitors

Screened complex karyotype cell lines for IC50, doubling time, in vivo growth, induction of autophagy

Performed autophagy-related gene array analysis (5 genes identified)

Specific aim 3 Performed growth and

clonigenicity assays Moved lab to OSU

Project 2Identification of therapeutic windows for NF1-related

malignant peripheral nerve sheath tumor

Co-Leaders Yuan Zhu, PhD (CNMC) Laurence Baker, DO (UM)

Specific Aims Aim 1: To investigate the cell-of-origin of PNST and

clonal relationship between plexiform neurofibroma and MPNST in humans

Aim 2: To determine phenotypic consequences of Erk/MAPK pathway inhibition during initiation phases of PNST development

Aim 3: To define a subset of NF1-associated MPNSTs responsive to MEKi

Project 2 Update

Aim 1: (1) IRB for the German site was approved. (2) All the methodologies including cell cultures for human Schwann cells and melanocytes as well as targeted NF1 sequencing were established. IRB for US sites are completed.

Aim 2: Multiple treatment protocols (n = 4) using an MEK inhibitor (MEKi from Pfizer) were employed on a mouse model for benign plexiform neurofibroma (PNF). Preliminary data were encouraging. This aim is expected to complete within a year. Parallel studies with rapamycin treatment were also conducted. The results were very promising and would be a nice comparison to human Rapa and MEKi clinical trials.

Aim 3: MEKi treatment showed no effect on established MPNSTs. Current efforts are to prepare for a manuscript to publish the new MPNST mouse models.

Project 3Investigating G-protein coupled receptors (GPCRs) as biomarkers of aggressive disease and novel therapeutic targets in Ewing sarcoma

Co-Leaders Elizabeth R. Lawlor, MD, PhD (UM) Rashmi Chugh, MD (UM)

Specific Aims Aim 1: Determine if C-X-C chemokine receptor type 4

(CXCR4)-mediated activation of RHO promotes invasion and metastasis

Aim 2: Evaluate the efficacy of Rho/MKL transcriptional antagonists as inhibitors of EFT cell invasion and metastasis

Aim 3: Validate the utility of GPCRs as biomarkers of aggressive disease in EFT

Two Manuscripts in preparation

Aim 1: CXCR4 expression in Ewing sarcoma is highly variable, dynamic and is induced by stress. CXCR4 promotes SDF1α (CXCL12)-dependent invasion via activation of Rac/Cdc42

CXCR4

CXCR4

+ +/- -

CXCR4 is reversibly induced by growth-factor deprivation (shown), hypoxia and growth-restriction.

AMD3100 blocks CXCR4- mediated invasion

Aim 2: Inhibition of MKL2 restores the actin cytoskeleton and blocks cell invasion

ControlInvasiveActive Rac/Cdc42

MKL-inhibitor Rx Non-invasiveInactive Rac/Cdc42

Inter- and intra-tumor heterogeneity of CXCR4 in primary tumors

Control

MKL-inhibitor Rx

Project 4Development of quantitative imaging biomarkers for

assessing response to sarcoma therapy

Co-Leaders Jeffrey Yap, PhD (Utah) Lawrence Schwartz, MD (Columbia)

Specific Aims Aim 1: Clinical validation of perfusion and diffusion MRI

imaging Aim 2: Development and preclinical validation of

imaging biomarkers for apoptosis Aim 3: Development and preclinical validation of

imaging biomarkers for angiogenesis

Project 4 Update

MRI acquisition protocols tested in healthy subjects for future clinical trial (Columbia, Utah, Michigan, Ohio State)

Preliminary mouse imaging studies performed using novel probes for angiogenesis ([89Zr]bevacizumab) and apoptosis ([18F]WC-II-89)

Future studies using 18F-fluciclatide and 124I-Diannexin are being developed

Developmental Research Program Grants

Recipient Project

Z. Duan Dissecting Cdk11 Kinase Pathway in Osteosarcoma

J. FletcherA. Marino-Enriquez

Biomarkers for Dystrophin-deficiency in Sarcoma

D. KirschR. Dodd

Translating Mouse Model Discoveries into Metastatic Biomarkers

S. LessnickS. Sharma

Epigenetic Targeting of LSD1 for Ewing Sarcoma

K. LiuR. Pollock

Combined Radiosensitization and Tcell Immunotherapy

D. LangenauC. Keller

GSK3-beta inhibitors for relapsed rhabdomyosarcoma

SARC Sarcoma SPORE

Career Development Awards

To identify and support talented new researchers in the area of sarcoma translational research

To enforce adequate protected time of clinically oriented young investigators to facilitate the development of independent translational researchers

Developmental Research Program

To identify and support talented researchers in the area of sarcoma translational research

To promote interdisciplinary research and move basic research findings from the laboratory to clinical settings

2014 Funding Opportunities for CDA and DRP Letter of intent due January 3, 2014 Details at SARC website