The 2014 program for the Connective Tissue Oncology Society (ctos.org) is online now. You may need to copy this link into your browser:
http://ctos.org/PDFs/CTOS%202014%20AM%2 ... tronic.pdf
OR
http://tinyurl.com/oxgychw
You can search for name (Alveolar soft part sarcoma) to see the studies that are relevant to us. I found several and will copy/past them later here.
CTOS 2014 meetings program
Re: CTOS 2014 meetings programm
Poster 168
PROTEOMICS IDENTIFIED OVEREXPRESSION OF SET NUCLEAR ONCOGENE PRODUCTS AND POSSIBLE THERAPEUTIC TARGET IN ALVEOLAR SOFT PART SARCOMA
Daisuke Kubota1; Kenta Mukaihara2; Yoshiyuki Suehara, MD, PhD2; Akihiko Yoshida4; Kazuo Kaneko2; Akira Kawai1; Tadashi Kondo3
1Division of Musculoskeletal Oncology, National Cancer Center Hospital, Tokyo, Japan; 2Department of Orthopedic Surgery, Juntendo University School of Medicine, Tokyo, Japan; 3Division of Pharmacoproteomics, National Cancer Center Reseasrch Institute, Tokyo, Japan; 4Pathology and Clinical Laboratory Division, National Cancer Center Hospital, Tokyo, Japan
Objective: Alveolar soft part sarcoma (ASPS) is an exceedingly rare sarcoma refractory to standard chemotherapy. Although several molecular targeting drugs have been applied for ASPS, their clinical significance has not yet been established, and novel therapeutic strategies have long been required. The aim of this study was to identify proteins aberrantly regulated in ASPS and to clarify their clinical significance.
Methods: Protein expression profiling of tumor and nontumor tissues from 12 ASPS patients was performed by 2-D difference gel electrophoresis and mass spectrometry.
Results: We found that the expression of 145 proteins differed significantly. Among them, further investigation was focused on the SET protein, which has multifunctional roles in cancers. Immunohistochemistry confirmed overexpression of SET in all 15 ASPS cases examined. Gene silencing of SET significantly decreased cell proliferation, invasion, and migration against a background of induced apoptosis. SET is known to be an inhibitor of phosphatase 2A (PP2A), which functions as a tumor suppressor by inhibiting the signal transduction pathway and inducing apoptosis. We found that a PP2A activator, FYN720, decreased cell proliferation through apoptosis.
Conclusion: Our findings may suggest the possible contribution of SET to the tumor progression and the utility of FYN720 for treatment of ASPS.
PROTEOMICS IDENTIFIED OVEREXPRESSION OF SET NUCLEAR ONCOGENE PRODUCTS AND POSSIBLE THERAPEUTIC TARGET IN ALVEOLAR SOFT PART SARCOMA
Daisuke Kubota1; Kenta Mukaihara2; Yoshiyuki Suehara, MD, PhD2; Akihiko Yoshida4; Kazuo Kaneko2; Akira Kawai1; Tadashi Kondo3
1Division of Musculoskeletal Oncology, National Cancer Center Hospital, Tokyo, Japan; 2Department of Orthopedic Surgery, Juntendo University School of Medicine, Tokyo, Japan; 3Division of Pharmacoproteomics, National Cancer Center Reseasrch Institute, Tokyo, Japan; 4Pathology and Clinical Laboratory Division, National Cancer Center Hospital, Tokyo, Japan
Objective: Alveolar soft part sarcoma (ASPS) is an exceedingly rare sarcoma refractory to standard chemotherapy. Although several molecular targeting drugs have been applied for ASPS, their clinical significance has not yet been established, and novel therapeutic strategies have long been required. The aim of this study was to identify proteins aberrantly regulated in ASPS and to clarify their clinical significance.
Methods: Protein expression profiling of tumor and nontumor tissues from 12 ASPS patients was performed by 2-D difference gel electrophoresis and mass spectrometry.
Results: We found that the expression of 145 proteins differed significantly. Among them, further investigation was focused on the SET protein, which has multifunctional roles in cancers. Immunohistochemistry confirmed overexpression of SET in all 15 ASPS cases examined. Gene silencing of SET significantly decreased cell proliferation, invasion, and migration against a background of induced apoptosis. SET is known to be an inhibitor of phosphatase 2A (PP2A), which functions as a tumor suppressor by inhibiting the signal transduction pathway and inducing apoptosis. We found that a PP2A activator, FYN720, decreased cell proliferation through apoptosis.
Conclusion: Our findings may suggest the possible contribution of SET to the tumor progression and the utility of FYN720 for treatment of ASPS.
Olga
Re: CTOS 2014 meetings programm
Poster 157
PRECLINICAL ACTIVITY OF KPT-330, AN INHIBITOR OF XPO1/CRM1, IN SEVERAL MODELS OF SARCOMA SUBTYPES
Robert Nakayama, MD, PhD1; Yi-Xiang Zhang1;
Jeffrey Czaplinski2; Ewa Sicinska2; Stephen Remillard1;
George D. Demetri1; Andrew J. Wagner1
1Ludwig Center at Dana-Farber/Harvard and Center for Sarcoma and Bone Oncology, Department of Medical Oncology, Harvard Medical School, Boston, MA, USA; 2Center for Molecular Oncologic Pathology, Dana-Farber Cancer Institute, Boston, MA, USA
Objective: XPO1/CRM1 is a nuclear export receptor involved in active transport of cargo proteins bearing nuclear export signals, including tumor suppressors (e.g. p53) and cell cycle regulators (e.g. p21). Selective inhibitors of nuclear export (SINEs) that link irreversibly to XPO1/CRM1 and block binding to cargo proteins are in clinical development as anticancer therapeutics. Prior preclinical and clinical studies have demonstrated activity primarily in hematologic malignancies as well as certain solid tumors. In this study, we evaluated the efficacy of an orally bioavailable SINE, KPT-330, in several preclinical models of various sarcoma subtypes.
Methods: The efficacy of KPT-330 was investigated in vitro and in vivo using 16 cell lines and 10 sarcoma xenograft models including GIST, liposarcoma (LPS), leiomyosarcoma, rhabdomyosarcoma, alveolar soft part sarcoma (ASPS), and undifferentiated sarcomas. Following exposure of cell lines to KPT-330, effects on cell viability, cell cycles, and RNA and protein expression were determined. In sarcoma xenograft studies, KPT-330 was administered twice a week by oral gavage. The size of subcutaneously implanted tumors was evaluated by measuring the long and short diameters.
Results: Most sarcoma cell lines were sensitive to KPT-330 with IC50s ranging from 28.8 nM to 218.2 nM (median: 67.6 nM). The ASPS cell lines were exceptionally resistant to KPT-330 with IC50 greater than 2 μM. In sarcoma xenograft studies, KPT-330 suppressed tumor growth, including ASPS models that were resistant in vitro. Cell lines from several sarcoma subtypes with defined molecular backgrounds, such as dedifferentiated LPS with MDM2 amplification and GIST with KIT mutations, were treated with KPT-330 to investigate its mechanism of action. KPT-330 induced G1-arrest and apoptosis in LPS cell lines irrespective of p53 expression or mutation status. KPT-330 increased p53 and p21 expression at the protein but not RNA level, indicating a post-transcriptional effect. KPT-330 induced G1 arrest in GIST cells without attenuation in phosphorylation of KIT, AKT, or MAPK, unlike imatinib.
Conclusion: KPT-330 has potent in vitro and in vivo activity against a wide variety of sarcoma models. KPT-330 induced G1-arrest independent of known molecular mechanisms in GIST and LPS. The difference in sensitivity of in vitro and in vivo ASPS models suggests a potential stromal effect. These studies further justify the exploration of KPT-
PRECLINICAL ACTIVITY OF KPT-330, AN INHIBITOR OF XPO1/CRM1, IN SEVERAL MODELS OF SARCOMA SUBTYPES
Robert Nakayama, MD, PhD1; Yi-Xiang Zhang1;
Jeffrey Czaplinski2; Ewa Sicinska2; Stephen Remillard1;
George D. Demetri1; Andrew J. Wagner1
1Ludwig Center at Dana-Farber/Harvard and Center for Sarcoma and Bone Oncology, Department of Medical Oncology, Harvard Medical School, Boston, MA, USA; 2Center for Molecular Oncologic Pathology, Dana-Farber Cancer Institute, Boston, MA, USA
Objective: XPO1/CRM1 is a nuclear export receptor involved in active transport of cargo proteins bearing nuclear export signals, including tumor suppressors (e.g. p53) and cell cycle regulators (e.g. p21). Selective inhibitors of nuclear export (SINEs) that link irreversibly to XPO1/CRM1 and block binding to cargo proteins are in clinical development as anticancer therapeutics. Prior preclinical and clinical studies have demonstrated activity primarily in hematologic malignancies as well as certain solid tumors. In this study, we evaluated the efficacy of an orally bioavailable SINE, KPT-330, in several preclinical models of various sarcoma subtypes.
Methods: The efficacy of KPT-330 was investigated in vitro and in vivo using 16 cell lines and 10 sarcoma xenograft models including GIST, liposarcoma (LPS), leiomyosarcoma, rhabdomyosarcoma, alveolar soft part sarcoma (ASPS), and undifferentiated sarcomas. Following exposure of cell lines to KPT-330, effects on cell viability, cell cycles, and RNA and protein expression were determined. In sarcoma xenograft studies, KPT-330 was administered twice a week by oral gavage. The size of subcutaneously implanted tumors was evaluated by measuring the long and short diameters.
Results: Most sarcoma cell lines were sensitive to KPT-330 with IC50s ranging from 28.8 nM to 218.2 nM (median: 67.6 nM). The ASPS cell lines were exceptionally resistant to KPT-330 with IC50 greater than 2 μM. In sarcoma xenograft studies, KPT-330 suppressed tumor growth, including ASPS models that were resistant in vitro. Cell lines from several sarcoma subtypes with defined molecular backgrounds, such as dedifferentiated LPS with MDM2 amplification and GIST with KIT mutations, were treated with KPT-330 to investigate its mechanism of action. KPT-330 induced G1-arrest and apoptosis in LPS cell lines irrespective of p53 expression or mutation status. KPT-330 increased p53 and p21 expression at the protein but not RNA level, indicating a post-transcriptional effect. KPT-330 induced G1 arrest in GIST cells without attenuation in phosphorylation of KIT, AKT, or MAPK, unlike imatinib.
Conclusion: KPT-330 has potent in vitro and in vivo activity against a wide variety of sarcoma models. KPT-330 induced G1-arrest independent of known molecular mechanisms in GIST and LPS. The difference in sensitivity of in vitro and in vivo ASPS models suggests a potential stromal effect. These studies further justify the exploration of KPT-
Olga
Re: CTOS 2014 meetings programm
Poster 40
BRAIN METASTASES IN SARCOMA PATIENTS, THE ROYAL MARSDEN HOSPITAL EXPERIENCE 1998 TO 2013: OUTCOMES AND PROGNOSTIC VARIABLES
Thomas M. Richards, MBBS, BSc, MRCP, FRCR1; Attilla Kollar1; Robert Urwin1; Komel Khabra2;
Omar Al-Muderis1; Charlotte Benson1; Mark Linch1;
Ian Judson1; Aisha Miah, MRCP FRCR PhD1
1The Sarcoma Unit, The Royal Marsden Hospital NHS Foundation Trust, London, United Kingdom;
2The Statistics Unit, The Royal Marsden Hospital NHS Foundation Trust, London, United Kingdom
Objective: Brain metastases (BM) have a low incidence, ~5-6% in sarcoma patients. Few studies have investigated the demographics and impact of local treatments in this poor prognosis sub-group or assessed prognostic variables associated with outcome.
Methods: A prospective database, collected at The Royal Marsden Hospital (RMH) from 1998-2013, recorded all patients presenting with BM who have a diagnosis of soft-tissue or bone sarcoma. Patient records were retrospectively reviewed to assess demographics, treatment and survival outcomes. Sequential univariate-multivariate analyses were performed to assess 7 variables for an independent association with survival.
Results: Between 02/1998 and 02/2013 eighty-nine patients with a primary sarcoma were diagnosed with BM. At a median follow-up of 4.8 years, 3 (3.2%) patients were alive. Mean patient age (range) was 47 years (18.4-89) with 55 (61.8%) male. Most frequent histologies were leiomyosarcoma (LMS) and pleomorphic sarcoma with 14 patients (15.7%) in each category. Primary tumour site was the limb in 33 (37.1%) patients. Median time (range) from diagnosis to detection of BM was 15.3 months (0-167.1) with a median (IQR) of 2 (1-4) BM detected. Following BM diagnosis, 68 patients received active treatment with whole brain or stereotactic radiotherapy (RT) (n=51), surgery (S) (n=5) or combination (C) of RT and surgery (n=12). Median (95% C.I.) survival (months) after RT, S or C was 3.2 (2.5-3.8 ), 18.1 (0-43.3) and 18.6 (14.4-22.7) respectively compared to best supportive care (BSC) of 1.1 (0-2.3). Variables, independently associated (p<0.05) with survival (Hazard Ratio, 95% C.I.) were the presence of extra-cranial disease (4.14, 1.66-10.31), age at BM diagnosis (1.03, 1.01-1.04), LMS (0.22, 0.1-0.5), alveolar soft-part sarcoma histology (0.3, 0.1-0.89) and surgical excision of BM (0.18, 0.09-0.39).
Conclusion: Patients who received surgery or combination treatment for BM had improved survival compared to best supportive care. Five variables were found to be significantly associated with survival and may be useful prognostic markers, to determine benefit of intervention in this uncommon sarcoma sub-group.
BRAIN METASTASES IN SARCOMA PATIENTS, THE ROYAL MARSDEN HOSPITAL EXPERIENCE 1998 TO 2013: OUTCOMES AND PROGNOSTIC VARIABLES
Thomas M. Richards, MBBS, BSc, MRCP, FRCR1; Attilla Kollar1; Robert Urwin1; Komel Khabra2;
Omar Al-Muderis1; Charlotte Benson1; Mark Linch1;
Ian Judson1; Aisha Miah, MRCP FRCR PhD1
1The Sarcoma Unit, The Royal Marsden Hospital NHS Foundation Trust, London, United Kingdom;
2The Statistics Unit, The Royal Marsden Hospital NHS Foundation Trust, London, United Kingdom
Objective: Brain metastases (BM) have a low incidence, ~5-6% in sarcoma patients. Few studies have investigated the demographics and impact of local treatments in this poor prognosis sub-group or assessed prognostic variables associated with outcome.
Methods: A prospective database, collected at The Royal Marsden Hospital (RMH) from 1998-2013, recorded all patients presenting with BM who have a diagnosis of soft-tissue or bone sarcoma. Patient records were retrospectively reviewed to assess demographics, treatment and survival outcomes. Sequential univariate-multivariate analyses were performed to assess 7 variables for an independent association with survival.
Results: Between 02/1998 and 02/2013 eighty-nine patients with a primary sarcoma were diagnosed with BM. At a median follow-up of 4.8 years, 3 (3.2%) patients were alive. Mean patient age (range) was 47 years (18.4-89) with 55 (61.8%) male. Most frequent histologies were leiomyosarcoma (LMS) and pleomorphic sarcoma with 14 patients (15.7%) in each category. Primary tumour site was the limb in 33 (37.1%) patients. Median time (range) from diagnosis to detection of BM was 15.3 months (0-167.1) with a median (IQR) of 2 (1-4) BM detected. Following BM diagnosis, 68 patients received active treatment with whole brain or stereotactic radiotherapy (RT) (n=51), surgery (S) (n=5) or combination (C) of RT and surgery (n=12). Median (95% C.I.) survival (months) after RT, S or C was 3.2 (2.5-3.8 ), 18.1 (0-43.3) and 18.6 (14.4-22.7) respectively compared to best supportive care (BSC) of 1.1 (0-2.3). Variables, independently associated (p<0.05) with survival (Hazard Ratio, 95% C.I.) were the presence of extra-cranial disease (4.14, 1.66-10.31), age at BM diagnosis (1.03, 1.01-1.04), LMS (0.22, 0.1-0.5), alveolar soft-part sarcoma histology (0.3, 0.1-0.89) and surgical excision of BM (0.18, 0.09-0.39).
Conclusion: Patients who received surgery or combination treatment for BM had improved survival compared to best supportive care. Five variables were found to be significantly associated with survival and may be useful prognostic markers, to determine benefit of intervention in this uncommon sarcoma sub-group.
Olga
Re: CTOS 2014 meetings programm
Paper 055
MODELING ALVEOLAR SOFT PART SARCOMAGENESIS IN THE MOUSE: A ROLE FOR LACTATE IN THE TUMOR MICROENVIRONMENT
Matthew L. Goodwin, MD, PhD1; Huifeng Jin1;
Krystal M. Straessler2; Kyllie Smith-Fry1; Ju-Fen Zhu1; Michael J. Monument1; Allie Grossmann3;
R. Lor Randall1; Mario R. Capecchi2;
Kevin B. Jones, MD1
1Department of Orthopaedics, University of Utah, Huntsman Cancer Institute, Salt Lake City, UT, USA; 2Department of Human Genetics and Howard Hughes Medical Institute, University of Utah, Salt Lake City, UT, USA; 3Department of Pathology, University of Utah,
Salt Lake City, UT, USA
Objective: Alveolar soft part sarcoma (ASPS) is a deadly soft tissue malignancy of unknown origin that consistently associates with the fusion gene ASPSCR1-TFE3 (AT3). Given its poor prognosis and the paucity of models, we aimed to generate a genetically-driven mouse model, confirm its recapitulation of human ASPS, and test the conditions necessary for transformation.
Methods: The Rosa26-LSL-AT3 allele was generated by isolating the fusion gene transcript from human ASPS RNA and targeting the cDNA to the Rosa26 locus behind a floxed stop for conditional induction.
Results: Mice activating AT3 expression randomly across all tissues formed tumors that were histologically indistinguishable from human ASPS with complete penetrance by 3-6 months. RNA sequencing of mouse and human ASPS tumors demonstrated strong transcriptomic mimicry. The most consistently up-regulated genes were those affecting carbohydrate metabolism, cell division, and cell cycle. Anatomically, the mouse tumors formed only within the cranial vault, while human ASPSs form mostly in skeletal muscle, frequently metastasizing to the brain. The consistency of this location even with different drivers of Cre-recombinase and different host tissues within the cranium suggested that something unique to the microenvironment inside the cranial vault enhanced tumorigenesis. Both brain and muscle are known to demonstrate high lactate flux. Measured steady-state lactate concentrations in normal mice identified brain and eye (the host tissue compartments of all the AT3-driven tumors), as having the most elevated levels. Mct1 and Cd147, known to associate with lactate import, have been shown to be overexpressed in ASPS. Functionally, mouse tumors responded oxidatively to administered lactate, supporting a role for lactate as a fuel. Tumors demonstrated abundant Hif1α despite no hypoxia, suggesting that lactate might be stabilizing Hif1α. Lactate administered by daily isotonic intraperitoneal injections drove dramatic increases in proliferative index and vascular density compared to saline.
Conclusion: We have generated a mouse model that recapitulates human ASPS with one notable discrepancy: location of tumors in the cranial vault. Investigation of this departure from the clinical biology of human ASPS led to discovery of a role for lactate in the microenvironment, which helps to explain the predilection of human ASPS for its frequently arising in muscle and metastasizing to brain and cardiac tissue, all high in lactate.
MODELING ALVEOLAR SOFT PART SARCOMAGENESIS IN THE MOUSE: A ROLE FOR LACTATE IN THE TUMOR MICROENVIRONMENT
Matthew L. Goodwin, MD, PhD1; Huifeng Jin1;
Krystal M. Straessler2; Kyllie Smith-Fry1; Ju-Fen Zhu1; Michael J. Monument1; Allie Grossmann3;
R. Lor Randall1; Mario R. Capecchi2;
Kevin B. Jones, MD1
1Department of Orthopaedics, University of Utah, Huntsman Cancer Institute, Salt Lake City, UT, USA; 2Department of Human Genetics and Howard Hughes Medical Institute, University of Utah, Salt Lake City, UT, USA; 3Department of Pathology, University of Utah,
Salt Lake City, UT, USA
Objective: Alveolar soft part sarcoma (ASPS) is a deadly soft tissue malignancy of unknown origin that consistently associates with the fusion gene ASPSCR1-TFE3 (AT3). Given its poor prognosis and the paucity of models, we aimed to generate a genetically-driven mouse model, confirm its recapitulation of human ASPS, and test the conditions necessary for transformation.
Methods: The Rosa26-LSL-AT3 allele was generated by isolating the fusion gene transcript from human ASPS RNA and targeting the cDNA to the Rosa26 locus behind a floxed stop for conditional induction.
Results: Mice activating AT3 expression randomly across all tissues formed tumors that were histologically indistinguishable from human ASPS with complete penetrance by 3-6 months. RNA sequencing of mouse and human ASPS tumors demonstrated strong transcriptomic mimicry. The most consistently up-regulated genes were those affecting carbohydrate metabolism, cell division, and cell cycle. Anatomically, the mouse tumors formed only within the cranial vault, while human ASPSs form mostly in skeletal muscle, frequently metastasizing to the brain. The consistency of this location even with different drivers of Cre-recombinase and different host tissues within the cranium suggested that something unique to the microenvironment inside the cranial vault enhanced tumorigenesis. Both brain and muscle are known to demonstrate high lactate flux. Measured steady-state lactate concentrations in normal mice identified brain and eye (the host tissue compartments of all the AT3-driven tumors), as having the most elevated levels. Mct1 and Cd147, known to associate with lactate import, have been shown to be overexpressed in ASPS. Functionally, mouse tumors responded oxidatively to administered lactate, supporting a role for lactate as a fuel. Tumors demonstrated abundant Hif1α despite no hypoxia, suggesting that lactate might be stabilizing Hif1α. Lactate administered by daily isotonic intraperitoneal injections drove dramatic increases in proliferative index and vascular density compared to saline.
Conclusion: We have generated a mouse model that recapitulates human ASPS with one notable discrepancy: location of tumors in the cranial vault. Investigation of this departure from the clinical biology of human ASPS led to discovery of a role for lactate in the microenvironment, which helps to explain the predilection of human ASPS for its frequently arising in muscle and metastasizing to brain and cardiac tissue, all high in lactate.
Olga