bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3096: Mapping lysine acetyltransferase-metabolite interactions in complex proteomes: a window into the metabolic regulation of epigenetic signaling
Authors: Meier Jordan L.
Published: July 2016

Substantial evidence indicates that chromatin modifiers are sensitive to changes in the cellular concentration of metabolites and cofactors. These findings potentially connect metabolism and gene expression, and have major implications for the regulation of cell signaling in cancer. However, missing from many of these studies has been the direct identification of the specific chromatin modifiers that sense change in metabolic state. Their identification is critical to our basic understanding of how metabolism regulates epigenetically-driven phenotypes, and may inform novel strategies to combat pathologies characterized by aberrant metabolism, including cancer. Towards this goal, here we report a chemical proteomic strategy to globally map chromatin modifier-metabolite interactions in complex proteomes. Our initial efforts have focused on lysine acetyltransferases (KATs), a central class of chromatin modifiers responsible for histone acetylation. First, we validate an optimized chemical proteomic method and demonstrate its ability to globally profile members of the KAT enzyme superfamily at endogenous expression levels. Then we apply this approach to rapidly profile KAT-ligand interactions, and identify novel candidate metabolic regulators of protein acetylation, including acyl-CoAs cofactors and metabolites, as well as new metabolically-sensitive KAT enzymes. By defining the molecular interactions linking metabolism and gene expression, these studies have the potential to illuminate our understanding of biology and inspire new therapeutic strategies targeting acetylation in disease.

Citation Format: Jordan L. Meier. Mapping lysine acetyltransferase-metabolite interactions in complex proteomes: a window into the metabolic regulation of epigenetic signaling. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3096.
Abstract 31: Metabolic reprogramming in ovarian cancer
Authors: HA JI HEE , Radhakrishnan Rangasudhagar , Ward Jeremy D. , Jayaraman Muralidharan , Dhanasekaran Danny N.
Published: July 2016

Ovarian cancer is currently the most lethal gynecologic malignancy, with no new significant changes in treatment options for these patients in the last 30 years. Importantly, ovarian cancer patients have increased levels of lysophosphatidic acid (LPA), a bioactive phospholipid in ascites and serum that has been linked to driving oncogenesis and progression of ovarian cancer. Accumulating evidence has implicated Hypoxia-inducible factor-1α (HIF-1α) as a critical mediator of the glycolytic shift observed in cancer cells. However, the precise biological role of LPA in regulating HIF-1α-mediated glycolytic shift is largely unknown. Therefore, we were interested in examining the potential role of LPA in promoting metabolic reprogramming in ovarian cancer via HIF-1α signaling. In this study, we identified a novel mechanism by which LPA upregulates HIF-1α expression via Gαi2 (the gip2 oncogene) in human ovarian cancer cells. This study demonstrates that LPA induces a glycolytic shift via LPA-mediated activation of Gαi2, which causes a subsequent upregulation of HIF-1α in ovarian cancer cells. Additionally, we show that LPA-signaling via Gαi2 induces an increase in the expression of Hexokinase-2 (HK2) and Glucose transporter 1 (GLUT1), which are known targets of HIF-1α. We also demonstrate that LPA induces an increase in extracellular acidification rate (ECAR) in a dose dependent manner in both ovarian cancer cell lines and in patient-derived cells taken from the ascites fluid of ovarian cancer patients using an XFe96 analyzer. Furthermore, we found that inhibition of Rac signaling caused a reduction in LPA-induced ECAR, identifying Rac as a critical downstream component of Gαi2-medidated increase of ECAR. Moreover, using NAC, an inhibitor of redox signaling, we found that this caused a decrease in LPA-induced ECAR in SKOV3-ip cells, indicating that inhibition of redox signaling abolishes LPA-induced ECAR in ovarian cancer cells. Similarly, the use of EUK-134, a scavenger of superoxide and H2O2, also decreased LPA-induced increase in ECAR. Altogether, these results indicate that LPA regulates glycolysis through redox signaling via a Gαi2-Rac-HIF1α-dependent signaling pathway. Most importantly, the Gαi2-Rac-dependent pathway identified in this study more than likely serves as a potential driver of HIF-1α-mediated metabolic changes in ovarian cancer cells and represents a potential target for therapy in these patients.

Citation Format: JI HEE HA, Rangasudhagar Radhakrishnan, Jeremy D. Ward, Muralidharan Jayaraman, Danny N. Dhanasekaran. Metabolic reprogramming in ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 31.
Abstract 3104: Stapled peptide inhibitors of the estrogen receptor/steroid receptor coactivator interaction
Authors: Moore Terry W. , Speltz Thomas E. , Fanning Sean W. , Mayne Christopher G. , Frasor Jonna , Greene Geoffrey L. , Tajkhorshid Emad
Published: July 2016

Despite undeniable successes, one of the major unsolved problems in estrogen receptor-positive breast cancer therapy is resistance to endocrine therapy. One-third of breast cancer patients who are given tamoxifen will develop recurrent cancer within 15 years. Importantly, the estrogen receptor is still present and active in 80-85% of these recurrent cases, but it is no longer sensitive to current therapies, so new ways of targeting the estrogen receptor are needed. Because selective estrogen receptor modulators indirectly block estrogen receptor/coactivator interactions, our hypothesis is that directly blocking the estrogen receptor/coactivator interaction will provide a more robust blockade that will antagonize forms of estrogen receptor that are active in both wild-type and some endocrine-resistant breast cancers.

As our starting point for inhibiting estrogen receptor/coactivator interactions, we have chosen to use a class of compounds that mimic α-helices: “stapled peptides.” These non-natural, synthetic peptides have a key alkene bond that resembles a staple. Typically, peptides make poor drugs because they are rapidly hydrolyzed by proteases and are poorly cell-permeable. Because of their non-natural linkage, stapled peptides are poor substrates for proteases, so they are metabolically stable. Some stapled peptides are also cell-penetrant. This effect is highly sequence-specific, but it seems to be aided when peptides have a relatively high formal positive charge. Stapled peptides have been used to inhibit a variety of important protein-protein interactions in cell culture and animal models. In this work, we have created a library of stapled peptides that inhibit the interaction of the estrogen receptor with the steroid receptor coactivator in vitro. The best peptides show nanomolar IC50 values in a time-resolved fluorescence resonance energy transfer assay. They show high helical content according to circular dichroism studies. We have solved x-ray crystal structures of these molecules bound to the estrogen receptor mutant Y537S, which demonstrate their binding mode, and these studies are in agreement with molecular dynamics simulations of these molecules bound to the estrogen receptor ligand binding domain.

Citation Format: Terry W. Moore, Thomas E. Speltz, Sean W. Fanning, Christopher G. Mayne, Jonna Frasor, Geoffrey L. Greene, Emad Tajkhorshid. Stapled peptide inhibitors of the estrogen receptor/steroid receptor coactivator interaction. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3104.
Abstract 3110: MASH1 expression in high grade astrocytoma may demonstrate a potential model for prognosis that may lead to a molecular target for future therapy
Authors: Tacha David E. , Tyrrell Jillian , Lobo Margaret
Published: July 2016

Introduction:

A large U.S. study showed that 25% of patients diagnosed with grade 3 astrocytoma survived for 5 years or more after diagnosis. Unfortunately, the outlook is even worse for patients diagnosed with GBM, as most patients live for less than a year. Achaete-scute complex homolog 1 (ASCL1, MASH1) is a proneural transcription factor that has an essential role in the formation of multiple neuronal lineages. Recent studies have shown that the upregulation of MASH1 and inhibition of the Notch signaling pathway characterize progressive astrocytoma. Proneural proteins have been shown to regulate stem-cell proliferation and differentiation. The convergence with oncogenic pathways combined with the overexpression in glioblastoma has been shown to correlate with poor clinical outcome and has identified these transcription factors as promising therapeutic targets. Recently, a MASH1 mouse monoclonal antibody has been developed. Most studies have shown MASH1 to be a specific marker of neuroendocrine carcinomas; however, only one immunohistochemical study on astrocytoma using a polyclonal antibody has been reported. This study examined MASH1 expression in grade 1-4 astrocytoma cases.

Materials and Methods:

Formalin-fixed paraffin-embedded tissue microarrays consisting of samples of normal brain and astrocytomas [grade 1 (n = 22), grade 2 (n = 34), grade 3 (n = 16) and glioblastoma (n = 35)] were evaluated by immunohistochemical analysis using a monoclonal MASH1 antibody. A value of >1% staining was scored as positive.

Results:

MASH1 stained 61.7% (66/107) of astrocytomas (grades 1-4). In grade 1 astrocytoma, MASH1 was expressed in 31.8% versus 85.3% in grade 2, 68.7% in grade 3 and 54.3% in grade 4 (p ∼ 0.0028).

Conclusion:

MASH1 is highly expressed in astrocytoma. There was a variable and progressive increase of MASH1 expression in grade 1 versus grade 2-4 astrocytoma; and thus, may demonstrate a potential model for prognosis that may lead to future target therapeutic strategies.

Citation Format: David E. Tacha, Jillian Tyrrell, Margaret Lobo. MASH1 expression in high grade astrocytoma may demonstrate a potential model for prognosis that may lead to a molecular target for future therapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3110.
Abstract 3117: Pleiotrophin is a marker of poor prognosis in Middle Eastern papillary thyroid carcinoma
Authors: Beg Shaham , Ahmed Maqbool , Hussein Azhar , Bu Rong , Qadri Zeeshan , AlSobhi Saif , Dayel Fouad Al , Siraj Abdul K. , Al-Kuraya Khawla S.
Published: July 2016

Pleiotrophin (PTN) is a heparin binding growth factor known to have role in neuronal development. It is highly expressed in embryo but has a very limited expression in adult tissues. PTN is considered a proto-oncogene and has been hypothesized to play role in oncogenesis as its expression is found to be increased in many different cancer subtypes. Its role in cell transformation, cell growth, survival, migration and angiogenesis has also been shown in various different types of cancers. The function of PTN is hypothesized to be carried out by its interaction with cell surface proteoglycans or binding to its selective cell surface receptor, protein tyrosine phosphatase receptor Z1 (PTPRZ1). The significance of role of PTN in pathogenesis of thyroid cancer has not been explored especially with the fact that papillary thyroid carcinoma (PTC) originating in this ethnic population is the second most common female malignancy, after breast. So in search for novel druggable molecular target we sought for PTN expression in a large cohort of Saudi PTC. We analysed PTN alteration in more than 1000 primary papillary thyroid carcinoma in a tissue microarray format with clinical follow up data. We found that PTN was overexpressed in 65.5% (658/1006) of PTC and was significantly associated with aggressive clinical parameters such as tall cell variant histological subtype (p = 0.0333), extrathyroidal extension (p = 0.0292), lymphovascular invasion (p = 0.0182) and large tumour size (p = 0.0160). Important significant molecular association was seen with PTPRZ-1 (p = 0.0316), Midkine (p = 0.0008) and pSTAT-3 (p<0.0001). Finally patients showing PTN overexpression have worse disease free survival compared to patients showing low or absent PTN expression (p = 0.0218). On multivariate analysis PTN is found to be an independent marker of poor prognosis taking into consideration parameters such as Age, Stage, Lymph node status and Extrathyroidal extension (p = 0.0430). This study highlights the importance of PTN as druggable molecular target which can be therapeutically exploited by its inhibitors in treatment of PTC.

Keywords: Papillary Thyroid Cancer; Pleiotrophin; Pleiotrophin Receptor

Citation Format: Shaham Beg, Maqbool Ahmed, Azhar Hussein, Rong Bu, Zeeshan Qadri, Saif AlSobhi, Fouad Al Dayel, Abdul K. Siraj, Khawla S. Al-Kuraya. Pleiotrophin is a marker of poor prognosis in Middle Eastern papillary thyroid carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3117.
Abstract 3125: KCNQ1 expression is a strong prognostic biomarker for disease recurrence in stage II and III colon cancer
Authors: den Uil Sjoerd H. , Coupe Veerle M.H. , Linnekamp Janneke F. , van den Broek Evert , Goos Jeroen A.C.M. , Delis-van Diemen Pien M. , Belt Eric J.T. , van Grieken Nicole C.T. , Scott Patricia M. , Vermeulen Louis , Medema Jan Paul , Bril Herman , Stockmann Hein B.A.C. , Cormier Robert T. , Meijer Gerrit A. , Fijneman Remond J.
Published: July 2016

Background: Colorectal cancer (CRC) is the third most common cancer worldwide. Accurately identifying stage II CRC patients at high risk of recurrence and stage III patients at low risk of recurrence are key unmet clinical needs. We previously identified KCNQ1 as a tumour suppressor gene of which loss of expression was associated with poor survival in patients with CRC liver metastases. The present study aimed to examine the prognostic value of KCNQ1 in stage II and III colon cancer patients.

Methods: KCNQ1 mRNA expression was assessed in 90 stage II colon cancer patients (AMC-AJCCII-90) using microarray gene expression data. KCNQ1 protein expression was evaluated by immuno-histochemistry on tissue microarrays of 386 stage II and III colon cancer patients.

Results: Low KCNQ1 mRNA expression in microsatellite stable (MSS) stage II colon cancers was associated with poor disease free survival (DFS) (HR 3.35; 95% CI 1.16-9.66; p<0.05). Loss of KCNQ1 protein expression from epithelial cells was strongly associated with poor DFS in MSS stage II (HR 3.82; 95% CI 2.04-7.14; p<0.0001), MSS stage III (HR 2.93; 95% CI 1.70-5.02; p = 0.0001) and MSI stage III colon cancers (HR 5.06; 95% CI 1.07-23.89; p<0.05). Multivariate analysis demonstrated KCNQ1 to have independent prognostic value in addition to established clinicopathological parameters such as angioinvasion, nodal stage en MSI-status.

Conclusion: We conclude that KCNQ1 is a strong prognostic biomarker for prediction of disease recurrence (HR∼4) and may aid stratification of patients with stage II MSS colon cancer and stage III MSI CRC for adjuvant chemotherapy. Because KCNQ1 protein expression is determined by immuno-histochemistry, this biomarker can be implemented in standard clinical care using existing workflows.

Citation Format: Sjoerd H. den Uil, Veerle M.H. Coupe, Janneke F. Linnekamp, Evert van den Broek, Jeroen A.C.M. Goos, Pien M. Delis-van Diemen, Eric J.T. Belt, Nicole C.T. van Grieken, Patricia M. Scott, Louis Vermeulen, Jan Paul Medema, Herman Bril, Hein B.A.C. Stockmann, Robert T. Cormier, Gerrit A. Meijer, Remond J. Fijneman. KCNQ1 expression is a strong prognostic biomarker for disease recurrence in stage II and III colon cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3125.
Abstract 3130: Adiponectin, Leptin, IGF1 and TNFα serum biomarker as noninvasive diagnosis of colon adenoma
Authors: Ashktorab Hassan , Soleimani Akbar , Nichols Alexandra , Sodhi Komal , Kannan Lakshmi , Adeyinka Laiyemo , Nouraie Mehdi , Brim Hassan
Published: July 2016

Background and Aim: The potential role of Adiponectin, Leptin, IGF1 and TNFα as biomarker in colon adenoma has not been studied. Therefore, we investigated the blood serum levels of these biomarkers in colorectal adenoma.

Method: The case-control study consisted of 198 African American patients with colon adenoma (cases) and 198 healthy individuals (controls) at Howard University Hospital. We used Elisa for biomarkers detection. Statistical analysis was performed by t-test and multivariate logistic regression.

Results: The differences in median leptin, Adiponectin, IGF1 and TNFα levels between control and case groups (6.7 vs.16.4), (11.3 vs.46.0), (4.5 vs.12.9) and (71.4 vs. 130.8) were statistically significant (p<0.05), respectively. In a multivariate model, the odds ratio (ORs) for Adiponectin, TNFα and IGF1 were 2.0 (95% CI = 1.6-2.5; P≤0.001), 1.5 (95% CI = 1.5- 2.0; P 0.004) and 1.6 (95% CI = 1.3-2.0; P≤ 0.001), respectively. There were positive correlations between serum Adiponectin and IGF1 concentrations with age (r = 0.17, P≤ 0.001 and r = 0.13, P = 0.009), also between TNFα, IGF1 and Leptin concentration with Body Mass Index (BMI) (r = 0.44, P≤ 0.001 and r = 0.11, P = 0.03; r = 0.48, P≤0.001), respectively. There was a negative correlation between Adiponectin concentration and BMI (r = -0.40, P≤0.001), respectively.

Conclusion: These data support the hypothesis that serum Adiponectin, IGF1 and TNFα are risk biomarkers for noninvasive detection of colorectal adenomas.

Citation Format: Hassan Ashktorab, Akbar Soleimani, Alexandra Nichols, Komal Sodhi, Lakshmi Kannan, Laiyemo Adeyinka, Mehdi Nouraie, Hassan Brim. Adiponectin, Leptin, IGF1 and TNFα serum biomarker as noninvasive diagnosis of colon adenoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3130.
Abstract 315: Potential role of androgen receptor in aromatase inhibitor resistance
Authors: Hunt Caleb , Lock Sarah E. , Shannon Cynthia E. , Varela-Gonzalez Nichole M. , Larsen Randolph K. , Schech Amanda J.
Published: July 2016

Clinical assessment of breast cancer tumors has shown that androgen receptor (AR) is expressed in more than 75% of cases, independent of molecular subtype. Specifically, clinical studies have shown that estrogen receptor (ER) and AR discordance correlates with endocrine resistance. Preclinical studies show that downregulation of ER results in a lack of estrogen mediated growth which is concurrent with upregulation of AR and its mediated gene targets. In addition, forced expression of AR was shown to induce resistance to tamoxifen, even when estrogen receptor was still expressed. Furthermore, ER positive breast cancer cells made resistant to tamoxifen express higher levels of AR and show sensitivity to abiraterone acetate. Together, these results implicate AR in endocrine resistance and suggest AR as a potential therapeutic target in a subset of these endocrine resistant cancers. The aim of this study was to better understand the role of AR in aromatase inhibitor (AI) resistance through the use of cell lines made resistant to AI through xenograft mouse modeling. In this study, MCF7 cells, MCF7 cells transfected with the human aromatase gene (MCF7Ca), and their aromatase inhibitor (AI) resistant counterparts, were analyzed for changes in AR and ER expression by qPCR and Western blot. Sensitivity of these cells to AR targeting agents was assessed by MTT assay. MCF7Ca cells displayed decreased expression of AR and increased expression of ER when compared to MCF7 cells. AI-resistant cell lines displayed modulation of ER and AR expression when compared to their AI-sensitive counterparts. Upregulation of AR was accompanied with increased AR transcriptional activity, as demonstrated by increased expression of PSA. Treatment of AI-resistant cells with AR targeting agents displayed increased sensitivity when compared to AI-sensitive cells. Together, these results suggest that upregulation of AR may contribute to AI-resistance, and that targeting AR in AI-resistant breast cancer may have therapeutic relevance.

Citation Format: Caleb Hunt, Sarah E. Lock, Cynthia E. Shannon, Nichole M. Varela-Gonzalez, Randolph K. Larsen, Amanda J. Schech. Potential role of androgen receptor in aromatase inhibitor resistance. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 315.
Abstract 3158: Prostate cancer derived biomarkers within platelets have the ability to predict therapeutic response in castration resistant patients
Authors: Tjon-Kon-Fat Lee-Ann , Lundholm Marie , Wurdinger Thomas , Thellenberg-Karlsson Camilla , Widmark Anders , Wikström Pernilla , Nilsson Jonas
Published: July 2016

Background: Novel therapies for castration resistant prostate cancer (CRPC) have been introduced in the clinic with possibilities for individualized treatment plans. Best practice of those expensive drugs requires predictive biomarker monitoring. In this article a novel platform for circulating biomarker analysis has been used to follow cancer-derived transcripts implicated in therapy resistance.

Method: The platelet population of blood samples and QRT-PCR were used to identify selected biomarkers in CRPC patient's prior chemo- or androgen synthesis (AS)-directed therapies. The association between biomarker statuses (positive vs. negative) and therapy response, progression-free survival (PFS) and overall survival (OS) was examined.

Results: 40 patients received either docetaxel (n = 17) or AS-directed (n = 23) therapy, with a therapy response rate of 29% respectively 48%. The cancer-associated biomarkers were present within the platelet fraction. Analyzing these biomarkers in the chemotherapy group were associated with a short OS (p = 0.025). In the AS-directed group, the biomarkers were associated with shorter PFS (p = 0.015), and with an increased risk of therapy failure (HR: 5,5; p = 0,023), as well as short OS (p = 0.012).

Conclusions: Analyzing circulating biomarkers in the platelet population enables us to predict who will benefit from AS-directed therapy with high accuracy, and platelet based analysis of cancer derived biomarkers may be used in treatment stratification of patients scheduled for AS-directed therapies.

Citation Format: Lee-Ann Tjon-Kon-Fat, Marie Lundholm, Thomas Wurdinger, Camilla Thellenberg-Karlsson, Anders Widmark, Pernilla Wikström, Jonas Nilsson. Prostate cancer derived biomarkers within platelets have the ability to predict therapeutic response in castration resistant patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3158.
Abstract 3161: Development and implementation of a non-invasive NGS-based diagnostic test for precision medicine in prostate cancer
Authors: Du Pan , Wang Amy , Cheng Tak , Kuo Winston Patrick
Published: July 2016

The growth and survival of prostate cancer tumors relies primarily on the functioning of the androgen receptor (AR) signaling pathway. Recent studies suggested that the presence of AR-V7, a c-terminal truncated form of AR, in circulating tumor cells (CTCs) of patients with castration-resistant prostate cancer (CRPC) is associated with inherent and/or acquired resistance to enzalutamide and abiraterone, the stand of care androgen deprivation therapies in prostate cancer. A series of data also indicate that AR-Vs (eg, including AR-V7, ARv567es, AR-T878A, AR-F876L, and Arv567es) may drive resistance in CRPC. Expression of AR-Vs has been shown to correlate with disease progression and shortened survival and AR-V7 is most abundant in CRPC specimens. Considering that truncated ARs with C-terminal loss (splice variants) lack a functional LBD and are constitutively active, C-terminal AR-directed therapies may not be effective, and novels agents are needed that target mutated ARs including AR-Vs. Circulating tumor cell (CTC)-based AR-V7 tests are currently being tested in the clinic. However, nearly half of the CRPC patients do not have enough CTCs for AR-V7 test, raising the request for a complementary, non-CTC platform to detect AR variants as well as other resistance markers in circulation.

Here we report the successful development of a non-invasive, next generation sequencing-based diagnostics platform that offers comprehensive profiling of genomic alterations in prostate cancer, including splicing variants (such as AR-V7), point mutation, copy number, and translocation using biofluid samples (blood and urine) from CRPC patients. To the best of our knowledge, this is the first non-invasive diagnostic test that measures genetic alterations especially AR-V7 in all patients with prostate cancer, regardless of their status of CTC enumeration. The successful development and clinical validation of this test has potential to enable precision medicine in prostate cancer.

Citation Format: Pan Du, Amy Wang, Tak Cheng, Winston Patrick Kuo. Development and implementation of a non-invasive NGS-based diagnostic test for precision medicine in prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3161.
Abstract 3162: Blood-based exosomal miRNAs as biomarkers for diagnosis and prognosis of clear cell renal cell cancer
Authors: Heinzelmann Joana , Baumgart Sophie , Hoelters Sebastian , Janssen Martin , Stöckle Michael , Junker Kerstin
Published: July 2016

Introduction & objectives: In previous studies we identified specific miRNA alterations in tumor tissues of clear cell renal cell cancer (ccRCC) with diagnostic and prognostic value relating to the presence of metastasis. For few years it has been known, that miRNAs are actively packed in exosomes which can be released into body fluids. Therefore, we hypothesize that in a simple blood based test we are able to use specific miRNA signatures as minimal invasive biomarkers for confirmation of the diagnosis and evaluation of the metastatic risk in ccRCC.

Materials & methods: Initially, exosomes were isolated from 1 ml serum of 10 ccRCC patients (metastatic and non metastatic tumors) and 10 healthy volunteers using appropriated exosome isolation kit protocol (Thermo Fisher Scientific). Quality of exosomes was proven using exosomal markers (CD63, CD81, Alix, Synthenin) and Nano Particle Tracking Analysis. Exosomal totalRNA was isolated using miRNeasy Mini Kit (Qiagen). For miRNA analyses total RNA was reverse transcribed using TaqMan Reverse Transcription Kit (Thermo Fisher Scientific), cDNA was preamplified using TaqMan® PreAmp Master Mix (Thermo Fisher Scientific) and real time PCR was performed using Gene Expression master mix (Thermo Fisher Scientific). Expression differences were calculated using REST Software and SPSS.

Results: We have shown that 1 ml serum is sufficient to analyze miRNA expression in exosomes. Exosomes isolated from serum exhibit high amount of exosomal markers and possess the typical exosomal size (30-120 nm). CcRCC serum samples are characterized by downregulation of several miRNAs (including miR-10b and miR451). Furthermore, specific miRNAs (including miR-30c) differentiate patients with or without metastases as well as healthy volunteers.

Conclusion: These initial data confirm our hypothesis that the tissue based miRNA signature could be used as biomarkers for detection of aggressive ccRCC analyzing exosomes from liquid biopsy. This minimal invasive blood based test is easy to handle in clinical practice and could be a powerful tool for early detection and monitoring of metastatic disease. To validate these results the expansion of the sample set is ongoing.

Citation Format: Joana Heinzelmann, Sophie Baumgart, Sebastian Hoelters, Martin Janssen, Michael Stöckle, Kerstin Junker. Blood-based exosomal miRNAs as biomarkers for diagnosis and prognosis of clear cell renal cell cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3162.
Abstract 3174: Colon cancer molecular subtypes: Concordance, effect on survival and selection of most representative preclinical models
Authors: Sztupinszki Zsófia , Győrffy Balázs
Published: July 2016

Introduction: Colon cancer is a genetically heterogeneous disease and there is an increasing need for more detailed classification of stage II and III patients. The NCCN guideline currently lists three FDA approved gene-expression based multi-gene prognostic tests for stage II-III colorectal cancer. However, it is undecided how many molecular subtype would most accurately describe these tumours. Here, we cross-validated known molecular subtypes for their concordance and capability to predict prognosis free survival. We utilized gene expression signatures to identify the most representative cell line model for each subtype.

Materials and methods: Using publicly available datasets from the GEO repository we established a database containing clinical and microarray data for 2,166 colon cancer patients. Gene expression measured on Affymetrix HG-U133A and HG-U133 Plus2.0 arrays was processed according to authors’ description for each classifier in R. We conducted a systematic review of the literature in order to identify gene-expression based classifiers. We assigned each sample for these classificators and then compared their potential to predict recurrence free survival.

Results: All together 22 different molecular subtypes were re-classified including the CCHS, CIN25, CMS, ColoGuideEx, ColoGuidePro, CRCassigner, MDA114, Meta163, ODXcolon, Oncodefender, TCA19, V7RHS classifiers as well as subtypes established by Budinska, Chang, DeSousa, Marisa, Merlos, Popovici, Schetter, Yuen, and Watanabe (first authors). When comparing the methods the most concordant classifiers were MDA114 and DeSousa (Cramer's V = 0.711). The highest efficacy to predict progression free survival in stage II-III patients was achieved by Yuen (p = 3.9e-05, HR = 2.9), Marisa (p = 2.6e-05, HR = 2.6) and Chang (p = 9e-09, HR = 2.35). We also assigned 61 colon cancer cell lines from four independent studies to the closest molecular subtype.

Conclusion: In summary, here we present a comprehensive analysis of human colon cancer molecular subtypes by cross-validation using independent transcriptomic datasets.

Citation Format: Zsófia Sztupinszki, Balázs Győrffy. Colon cancer molecular subtypes: Concordance, effect on survival and selection of most representative preclinical models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3174.
Abstract 3179: Germline deletion of FH in hereditary leiomyomatosis and renal cell carcinoma (HLRCC)
Authors: Vocke Cathy D. , Ricketts Christopher J. , Middelton Lindsay A. , Yang Youfeng , Linehan. W. Marston
Published: July 2016

Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is a familial cancer syndrome characterized by germline mutations in the fumarate hydratase (FH) gene that encodes the enzyme responsible for conversion of fumarate to malate in the Kreb's Cycle. HLRCC is associated with the development of cutaneous and uterine leiomyomas, and a highly aggressive form of type 2 papillary kidney cancer. HLRCC renal tumors can be extremely aggressive with patients demonstrating locally advanced or disseminated disease even when associated with small tumors 1-2 cm in size. Therefore, accurate identification of the “at risk” individuals is very important. In our experience, FH point mutations are identified in 90% of individuals that are clinically diagnosed with HLRCC, while in the remainder, FH may be inactivated by deletion or other causes. Several studies have reported complete deletions of the FH gene and a few have estimated the extent of these deletions and identified the additional genes that would be lost in conjunction with FH. Interestingly, families with complete FH deletion have demonstrated cutaneous and uterine leiomyomas but no families have yet been reported with evidence of kidney cancer. A single family has been identified that demonstrates a partial deletion resulting in the loss of exon 1, and in this case kidney cancer has identified within the family. In this study, patients with suspected HLRCC hereditary kidney cancer but no germline point mutation were enrolled on an NCI-IRB approved protocol and an Agilent custom high-definition comparative genomic hybridization (CGH) array was used to identify FH gene deletions. The assay identified complete germline deletion of a single copy of FH within seven suspected HLRCC patients deletion ranging from 249kb to 4.74Mb that in all cases resulted in a minimum addition loss of the KMO, OPN3, CHML, and WDR6 genes. All results were confirmed by CLIA approved tests and a further five new patients were identified by direct CLIA testing, including a novel partial deletion. In contrast to previous reports, seven out of twelve (58.3%) patients/families in our cohort demonstrated a personal or family history of HLRCC associated Type 2 papillary kidney cancer. In conclusion, complete or partial deletion of the FH gene is an infrequent but important cause of HLRCC and has no protective effect on the incidence of HLRCC associated type 2 papillary kidney cancer.

Citation Format: Cathy D. Vocke, Christopher J. Ricketts, Lindsay A. Middelton, Youfeng Yang, W. Marston Linehan. Germline deletion of FH in hereditary leiomyomatosis and renal cell carcinoma (HLRCC). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3179.
Abstract 3191: Molecular testing in lung cancer: Patient and caregiver experiences
Authors: King Jennifer C. , Rigney Maureen
Published: July 2016

Increasingly over the past decade, new targeted therapeutics and immunotherapies with associated companion diagnostics have been approved for the treatment of lung cancer. Lung cancer is often associated with elderly and lower socio-economic populations that may not have a high level of health literacy. As part of a comprehensive needs assessment survey of the lung cancer community, lung cancer patients and their caregivers were asked whether they or their loved ones had undergone molecular testing/biomarker testing as well as asked to report the history of oncologic drugs they had taken. Demographic information and additional cancer treatment history were also collected. Over 300 survey responses were collected. Interim analyses indicated that the majority (68%) of patients currently undergoing active treatment had received molecular testing to help determine their treatment plan. There was a clear improvement in testing rates over time with less than 30% of patients who had completed treatment reporting use of a molecular test. Importantly, there is still an educational gap that needs to be addressed as greater than 20% of patients and greater than 30% of loved ones, including the group who self-identified as the primary caregiver, reported that they did not know whether molecular testing had been performed. In addition, patients reported that they had not received EGFR mutation testing but had taken erlotinib, gefinitib, and/or afatinib. Final results will be presented as well as an analysis of factors that associate with lack of molecular testing. Taken together, these data indicate that there is a critical need for enhanced patient and caregiver education regarding molecular testing.

Citation Format: Jennifer C. King, Maureen Rigney. Molecular testing in lung cancer: Patient and caregiver experiences. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3191.
Abstract 3195: Synergistic effects of combined treatment with hAT-MSC.sTRAIL and panobinostat in brainstem glioma
Authors: Choi Seung Ah , Lee Chanhee , Kwak Pil Ae , Wang Kyu-Chang , Phi Ji Hoon , Lee Ji Yeoun , Chong Sangjoon , Kim Seung-Ki
Published: July 2016

Human adipose-derived mesenchymal stem cells expressing secreted form of the tumor necrosis factor-related apoptosis-inducing ligand (hAT-MSC.sTRAIL) have demonstrated therapeutic activity against various tumors. However, sTRAIL resistance remains a challenge in developing anticancer strategies. To solve this problem, many studies have tried to combine drugs to produce synergism or sensitize resistant cancer cells. Histone deacetylase inhibitors (HDACi) have been known to induce expression of TRAIL death receptors 4 and 5 (DR4/DR5). Herein, we evaluated the use of combined therapy of hAT-MSC.sTRAIL with HDAC inhibitor, panobinostat, in enhancing sensitivity to hAT-MSC.sTRAIL mediated apoptosis against the brainstem glioma.

A sTRAIL was introduced into the characterized hAT-MSCs using electroporation. To confirm appropriate treatment concentration of panobinostat to the glioblastoma cells, dose titration was tested using cell viability assay. The therapeutic effect of single or combination treatment against glioblastoma was evaluated using primary cultured glioblastoma cells and cell lines. Orthotopic xenograft brainstem glioma mouse model was established using engineered firefly luciferase expressing tumor cells for bioluminescence in vivo imaging.

Panobinostat induced anti-proliferative effects in dose and time-dependent manner (IC50 range, 0.05-0.2 μM) and effectively enhanced the expression of TRAIL DR4 and DR5, but not decoy receptors. Combined hAT-MSC.sTRAIL and panobinostat significantly decreased the tumor cell growth compared to each alone. Intriguingly, the combination treatment not only induced apoptosis but also autophagy. Using a preclinical brainstem mouse model, we confirmed that the combination of hAT-MSC.sTRAIL and panobinostat was safe and induced regression of tumor volume. Furthermore, the combination therapy prolonged the survival of brainstem glioma-bearing mice.

Our results suggested that combination therapy of panobinostat enhanced the anti-cancer effect of hAT-MSC.sTRAIL and represent potential therapeutic approach to the brainstem glioma.

Citation Format: Seung Ah Choi, Chanhee Lee, Pil Ae Kwak, Kyu-Chang Wang, Ji Hoon Phi, Ji Yeoun Lee, Sangjoon Chong, Seung-Ki Kim. Synergistic effects of combined treatment with hAT-MSC.sTRAIL and panobinostat in brainstem glioma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3195.
Abstract 3214: A fully human anti-TIM3 antibody with co-stimulatory activity
Authors: Gray J Dixon , Krapf Irina , Zhou Heyue , Kaufmann Gunnar
Published: July 2016

Targeting immune checkpoint molecules has been demonstrated to have significant benefit to patients with cancer. One such molecule is TIM3 whose expression is associated with a phenomenon referred to as T cell exhaustion. While it has been suggested that ligation of TIM3 provides a negative signal to the T cell, there is no obvious inhibitory signaling motif associated with the molecule. We have developed a fully human antibody to TIM3 which demonstrates potent co-stimulatory activity. T cells stimulated with anti-CD3 plus either anti-CD28 or anti-TIM3 show comparable levels of cell activation. Similar to co-stimulation with anti-CD28, the anti-TIM3 antibody promotes expression of CD25, interferon gamma, Tbet and granzyme B, amongst others. Thus, anti-TIM3 co-stimulation promotes T cells with a pro-inflammitory phenotype. With TIM3 being associated with exhausted T cells, co-stimulation with this antibody would be expected to specifically re-activate these exhausted T cells at the tumor site. Moreover, since some NK cells constitutively express TIM3, it would be expected that this antibody would activate these cells as well. Altogether, we have identified a novel anti-TIM3 antibody that is capable of targeting T cells at the tumor site and activating them.

Citation Format: J Dixon Gray, Irina Krapf, Heyue Zhou, Gunnar Kaufmann. A fully human anti-TIM3 antibody with co-stimulatory activity. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3214.
Abstract 322: Selective inhibition of MYC expression by a small molecule G-quadruplex stabilizer
Authors: Leon Elena C. , Simmons John K. , Calabrese David , DuBois Wendy , Felsenstein Kenneth , Saunders Lindsey B. , Zhang Shuling , Michalowski Aleksandra , Gonzalez Frank , Mock Beverly A. , Schneekloth John S.
Published: July 2016

MYC overexpression or amplification is commonly seen in a wide variety of tumor types, including multiple myeloma. Although MYC appears to be an important driver of the oncogenic phenotype, the helix-loop-helix topology, short half-life, and rapid replenishment of its encoded protein make it a particularly challenging drug target. However, the promoter region of the MYC gene contains a G-quadruplex (G4) DNA secondary structure that can be stabilized to block transcription. Recently, we reported the identification of a compound from a small molecule microarray screen that selectively binds the MYC G4, compared with other oncogenes that also contain G4s, and inhibits MYC mRNA and protein expression, leading to cell cycle arrest in human multiple myeloma cell lines (ACS Chem Biol. 2015). Here we describe the in vitro and in vivo evaluation of a more potent analog that retains the selectivity of the original compound for the MYC G4 sequence. After observing strong growth inhibitory activity of this new compound in a diverse panel of human cancer cell lines, we evaluated its pharmacokinetic and toxicity profiles in mice. Preliminary pharmacokinetic studies showed promising serum bioavailability and exposure properties when administered either intravenously or intraperitoneally. The compound was well tolerated in a month long, dose-escalation toxicity study in mice receiving daily administration of the compound. No adverse effects were observed during the study. In an assessment of short term in vivo activity, MYC expression was inhibited in multiple myeloma xenografts. More long-term studies to evaluate the anti-tumor activity of the compound are currently in progress. Thus, our data provide evidence that small molecule stabilization of the MYC G4 can drive transcriptional silencing of oncogenic MYC expression both in vitro and in vivo.

Citation Format: Elena C. Leon, John K. Simmons, David Calabrese, Wendy DuBois, Kenneth Felsenstein, Lindsey B. Saunders, Shuling Zhang, Aleksandra Michalowski, Frank Gonzalez, Beverly A. Mock, John S. Schneekloth. Selective inhibition of MYC expression by a small molecule G-quadruplex stabilizer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 322.
Abstract 3224: Blocking FSTL1 reprograms cancer-caused abnormal immunity
Authors: Kudo-Saito Chie , Toyoura Masayoshi , Shoya Yuji , Ishida Akiko , Kon Ryoko
Published: July 2016

Purposes:

The close connection between imbalanced immunity and tumor progression has been increasingly recognized, and a number of inhibitors for blocking immune inhibitory checkpoints have been established to improve the anti-tumor immunity in cancer patients. However, clinical responses are limited to a part of the treated patients. We previously identified FSTL1, which is significantly secreted from Snail+ metastatic tumor cells, as a key effector molecule in the cancer-caused immune suppression and dysfunction. FSTL1 generates and expands pluripotent mesenchymal stem cells (MSCs), which are able to generate various immunoregulatory cells and functionally impaired CD8-low T cells, and also to induce de novo tumor dissemination into bone marrow. FSTL1 also directly confers higher invasiveness on tumor cells (EMT induction). To prevent the FSTL1-caused vicious circulation for tumor progression, we attempted to establish anti-FSTL1 blocking mAbs.

Results:

We generated chicken/mouse chimeric anti-FSTL1 mAbs, which specifically recognized human and mouse FSTL1. The anti-FSTL1 mAb significantly suppressed tumor proliferation and invasion, MSC induction in mouse bone marrow cells, and Treg induction in human PBMCs in vitro as compared to the control group. When bone metastatic melanoma models (C57BL/6 mice implanted with snail-transduced B16-F10 melanoma cells both subcutaneously and intravenously) were intraperitoneally injected with the anti-FSTL1 mAb (10 mg/kg) on Day 5 (when tumor metastasis was already observed in bone marrow) and Day 10 after tumor implantation, the FSTL1-caused events were inhibited, and tumor-specific CD8+ T cells with higher CTL activities were generated, resulting in significant suppression of subcutaneous tumor growth and bone metastasis in the treated mice as compared to those of the control mice. At least in the same models, conventional immune checkpoint inhibitors including anti-CTLA4, anti-PD1, and anti-PDL1 mAbs failed to improve the anti-tumor immunity, and rather promoted bone metastasis because of no reduction of regulatory cells, particularly the MSCs in the treated mice.

Conclusions:

These results suggest that blocking FSTL1 reprograms and properly induces anti-tumor immunity by switching the tumor-supportive immune directivity to the active mode against cancer. Our anti-FSTL1 mAb having totally different molecular mechanisms from the conventional ones may be a promising therapeutics as a next generation of “immune checkpoint inhibitors” for radically correcting cancer-manipulated immunity in clinical settings.

Citation Format: Chie Kudo-Saito, Masayoshi Toyoura, Yuji Shoya, Akiko Ishida, Ryoko Kon. Blocking FSTL1 reprograms cancer-caused abnormal immunity. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3224.