bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4671: Dual mutational changes of ALK gene fusion combined with EGFR or K-ras oncogenes in lung adenocarcinomas
Authors: Lee Jieun , Hong Suk Hee , Jeon Eun Kyung , Kim Jung O. , Kim Seung Joon , Kim Young Kyoon , Park Jae Gil , Sung Sook-Whan , Kim Tae-Jung , Kang Jin Hyoung
Published: May 2015

Purpose

The mutations of the EGFR, K-ras or ALK oncogenes are closely associated with tumorigenesis and progression of lung adenocarcinoma. Their mutational changes are known to be mutually exclusive, but double oncogenic mutations of ALK combined with EGFR or K-ras gene are rarely found. We analyzed the clinical impacts of double oncogenic mutations on clinical characteristics and treatment outcome, according to EGFR, K-ras, ALK mutation or triple negative (EGFR, K-ras, ALK wild) in lung adenocarcinomas.

Patients and Methods

For one hundred ninety six lung cancer patients conformed histologically as adenocarcinoma, we examined activating EGFR (exons 18 and 21) and K-ras (exon 2 and 3) mutations by direct nucleotide sequencing and. ALK gene rearrangements by FISH using break-apart probe. Among 196 patients, 86 patients (43.9%) were triple negative adenocarcinoma, 73 patients (37.2%) presented with activating EGFR mutation, 17 patients (8.7%) with ALK and 13 patients (6.6%) with K-ras mutation. Median age of 17 patients harboring ALK rearrangement was 54 years (range 25∼79), approximately 7 years younger than the patient population harboring EGFR or K-ras mutation (62 and 61 years, respectively). Fifteen patients (88.2%) had stage III or IV and 9 patients (52.9%) were never smoker. Platinum doublet chemotherapy or EGFR TKIs was administered for 14 patients initially presented as stage IV disease or recurred as metastatic disease after local treatment. The median PFS of the patients receiving platinum doublet chemotherapy and EGFR TKIs as first-line treatment was 5.67 months (range 2.97-20.9) and 3.58 months (range 3.53-3.63), respectively. Of 17 ALK-positive patients, dual oncogenic mutations of ALK together with either EGFR or K-ras were identified in 7 patients (3.6%). Six patients (85.7%) receiving platinum-doublet or EGFR TKI as initial treatment. showed partial response, and 1 patient (14.3%) showed stable disease. These seven patients are all alive at present.

Conclusion

Taken together, our data suggest the existence of dual oncogenic mutations in lung adenocarcinomas is not infrequent. We are undergoing the experiment for detection for circulating tumor cells harboring ALK rearrangements, EGFR or K-ras mutation in these patients' peripheral blood.

Citation Format: Jieun Lee, Suk Hee Hong, Eun Kyung Jeon, Jung Oh Kim, Seung Joon Kim, Young Kyoon Kim, Jae Gil Park, Sook-Whan Sung, Tae-Jung Kim, Jin Hyoung Kang. Dual mutational changes of ALK gene fusion combined with EGFR or K-ras oncogenes in lung adenocarcinomas. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4671. doi:10.1158/1538-7445.AM2014-4671
Abstract 468: Cancer cells with G/G mdm2 SNP309 have compromised transcriptional elongation of p53 target genes
Authors: Rosso Melissa , Polotskaia Alla , Bargonetti Jill
Published: May 2015

MDM2 protein is often found over-expressed in cancers with wild-type p53. A single nucleotide polymorphism (T to G) near the mdm2 P2 promoter, mdm2 SNP309, leads to MDM2 overexpression. This polymorphism is associated with accelerated tumor formation, decreased sensitivity to DNA damage treatment and compromised p53 transcriptional activity. Two G/G SNP309 cell lines MANCA (Burkitt lymphoma) and A875 (melanoma) express wild-type p53 protein not readily degraded by MDM2. This wild-type p53 does not activate target genes even after DNA damage. During the blocked transcription, MDM2 and p53 co-localize at p53 response elements on chromatin. Therefore, we hypothesized that MDM2 localized with p53 on the chromatin was the contributing factor for the compromised p53 transcriptional activity. We explored the mechanism for compromised p53 transcriptional activity in these G/G SNP309 cancer cell lines. When compared to fully activated wild-type p53 in ML-1 cells treated with etoposide, MANCA and A875 cell lines had comparable recruitment of total and initiated RNA pol II at transcription start sites (TSS) for p21 and puma genes. This indicated that G/G SNP309 cells had functional transcription initiation at p53 target genes after DNA damage treatment. We next assessed epigenetic modifications associated with active transcriptional elongation. Using H3K36 trimethylation (H3K36me3) as a hallmark for active elongation, we observed that ML-1 cells had a substantial increase in H3K36me3 after etoposide treatment at p21 and puma TSS that was not seen in MANCA or A875 cell lines. This lack of H3K36me3 suggested an inhibition in early transcriptional elongation. To address the participation of MDM2 in the elongation block we created a stable MDM2 knockdown MANCA cell line. Interestingly, MDM2 knockdown drastically increased MDMX protein levels and moderately increased activation of p53 target genes implying MDMX played a redundant role for MDM2. We are currently investigating the roles of MDM2 and MDMX in blocking transcriptional elongation. Interestingly, we have found that treatment of MANCA cells with low dose Actinomycin D reactivates the p53 pathway and studies are in progress to determine if this is through activation of transcriptional elongation.

This work was supported by The Breast Cancer Research Foundation and The National Science Foundation (grant MCB-0744316) to J.B. M.R. was supported by the MBRS-RISE Program to Hunter College 3R25-GM060665.

Citation Format: Melissa Rosso, Alla Polotskaia, Jill Bargonetti. Cancer cells with G/G mdm2 SNP309 have compromised transcriptional elongation of p53 target genes. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 468. doi:10.1158/1538-7445.AM2014-468
Abstract 4682: Recurrence risk factors for advanced gastric cancer after D2 gastrectomy followed by adjuvant chemotherapy
Authors: Hwang Jun-Eul , Kim Min-Jee , Shim Hyun-Jeong , Bae Woo-Kyun , Cho Sang-Hee , Chung Ik-Joo
Published: May 2015

Background

Recent trials demonstrated that the adjuvant chemotherapy for stage II or III gastric cancer after curative D2 gastrectomy is effective for preventing recurrence. The objective of this study was to clarify the risk factors for recurrence in patients who received adjuvant chemotherapy after curative D2 gastrectomy.

Materials and methods

We retrospectively analyzed the factors predicting recurrence in 206 patients with stage II or III gastric cancer who received oral S-1 or 5-FU and cisplatin (FP) chemotherapy following curative D2 gastrectomy between April 2004 and December 2011.

Results

The median follow-up time after surgery was 2.93 years. A total of 136 patients received FP chemotherapy, and 70 patients received S-1. 3 year recurrence-free survival was 66.7%. In univariate analysis, gastroesophageal junction cancer, stage III, T3/T4 tumor, N2/N3 nodal status, and presence of both lymphovascular invasion (LVI) and perineural invasion (PNI) were associated with shorter recurrence-free survival. Moreover, Multivariate analysis showed male (HR 1.909; 95% CI 1.067-3.417; p=0.029), stage III (HR 3.884; 95% CI 1.741-8.662; p=0.001), and presence of both LVI and PNI (HR 2.293; 95% CI 1.367-3.846; p=0.002) were independent risk factors for recurrence.

Conclusion

Presence of both LVI and PNI is a strong recurrence risk factor for patients with stage II and stage III R0 resected gastric cancer who received adjuvant chemotherapy.

Citation Format: Jun-Eul Hwang, Min-Jee Kim, Hyun-Jeong Shim, Woo-Kyun Bae, Sang-Hee Cho, Ik-Joo Chung. Recurrence risk factors for advanced gastric cancer after D2 gastrectomy followed by adjuvant chemotherapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4682. doi:10.1158/1538-7445.AM2014-4682
Abstract 4687: The PI3K inhibitor GDC-0941 is synergistic with lapatinib, and mediates endocrine sensitivity in uterine papillary serous carcinoma
Authors: Levano Kelly S. , Rodriguez-Gabin Alicia , Goldberg Gary L. , Horwitz Susan B. , Hou June Y.
Published: May 2015

Objectives: Our objective is to determine the effect of PI3K activity in mediating ligand-independent estrogen signaling and endocrine sensitivity in Uterine Papillary Serous Carcinoma (UPSC).

Methods: Drug effect on cell proliferation was calculated via Calcusyn in patient-derived UPSC cell lines ARK 1 and 2. PI3K mutation was analyzed after laser capture microdissection of tumor DNA in 7 consecutive patients with UPSC. Protein expression, via Western Blot, was correlated prospectively with clinical parameters.

Results: Table 1 shows cells line baseline characteristics and IC50 to drugs. Fulvestrant, an ER antagonist, rendered the cells significantly more resistant to taxol in ARK1 and 2(p=0.035, 0.021 respectively). The concomitant decrease in pERS167 and pAKTS473 with Fulvestrant treatment is independent of baseline ER expression and PI3K mutation. In ARK2, a cell line that is ER-, disrupting AKT signaling via the PI3K inhibitor GDC-0941, or with the lapatinib is synergistic with Fulvestrant with Combination Index (CI)50 of 0.441 and 0.229, respectively. Independent of HER2 amplification or PI3K mutation, lapatinib and GDC-0941 exhibited synergistic cytotoxicity in both UPSC cell lines (CI50 of 0.577, ARK 1 and 0.233, ARK2). Finally, PI3K mutation and pAKTS473 expression was analyzed in 7 tumor samples. While none harbored PI3K mutation, patients who are chemotherapy resistant had significantly lower expression of baseline pAKTS473 in their tumor samples than those who are chemosensitive, similar to our cell line data.

Conclusions: PI3K pathway dysregulation, either via upstream erbB amplification, or downstream constitutive activation of AKT, may be important in mediating endocrine and taxol sensitivity in UPSC. pAKTS473 may be an important biomarker of drug sensitivity. The combination of PI3K inhibitor and lapatinib is synergistic and warrant further therapeutic investigation.

Baseline characteristics and drug sensitivities in UPSC cell linesARK1ARK2PI3KCA mutationfExon 9NoERα (WB)YesNopERα167 (WB)YesNopHER (WB)NoYesIC50 nM (mean ± STD)GDC-094193.1±5.9271±116.5LapatinibNo effect135.6±29.2FulvestrantNo effectNo effectTaxol4.6±1.97.8±5.6Cisplatin1103±65.72632.6±1797.9

Citation Format: Kelly S. Levano, Alicia Rodriguez-Gabin, Gary L. Goldberg, Susan B. Horwitz, June Y. Hou. The PI3K inhibitor GDC-0941 is synergistic with lapatinib, and mediates endocrine sensitivity in uterine papillary serous carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4687. doi:10.1158/1538-7445.AM2014-4687
Abstract 4689: Pre-clinical investigation of the wee1 inhibitor MK-1775 using pharmacodynamic and mechanistic markers in diverse cancer models in vivo
Authors: Wilsker Deborah F. , Marrero Allison M. , Hollingshead Melinda , Lawrence Scott M. , Chen Alice , Kummar Shivaani , Covey Joseph M. , Parchment Ralph E. , Kinders Robert J. , Tomaszewski Joseph E. , Doroshow James H.
Published: May 2015

Wee1 kinase is a critical regulator of the G2/M checkpoint by initiating inhibitory phosphorylation of the conserved tyrosine-15 residue of cdk1/cdc2. Wee1 also phosphorylates tyrosine 15 of cdk2 and plays a crucial role in maintaining genome integrity during S-phase. MK1775 (NSC 754352) is a small molecule selective inhibitor of Wee1 kinase that is currently under clinical evaluation. We have developed a quantitative immunofluorescence assay to measure inhibition of Wee1 by detection of pY15 of cyclin-dependent kinases. A full analysis of the pharmacodynamic (PD) effects of Wee1 inhibition by MK1775, as well as the downstream mechanistic consequences of Wee1 inhibition on DNA repair, apoptosis, and premature mitotic entry markers was performed using two xenograft models: A673 Ewing sarcoma and U87-MG glioblastoma. Four DNA repair markers (pNbs1, γH2AX, Rad51, and pATR) were examined as well as the mitotic marker pHistone H3. Advanced quantitative image analysis was performed using Definiens software to measure changes in PD markers by two approaches: total nuclear area measurements and foci per nucleus. Definiens software enables enhanced analysis of the markers by enumeration of the nuclei in the imaged xenograft tissues over an entire data set with high-content capacity. Our data demonstrate greater than 80% inhibition of pY15-cdk in vivo at the clinically relevant dose of 60 mg/kg (180 mg/m2) MK1775 in two xenograft models. In addition, γH2AX induction was observed after multiple doses of MK1775 as a single agent. A PD biomarker time course was determined for MK1775 and gemcitabine (NSC 750927) as single agents to develop a drug administration schedule for combination studies in a Ewing sarcoma model. The time point at which the DNA damage response peaked following administration of the maximum tolerated dose of gemcitabine in the mouse was determined. This study elucidates a broad profile of PD marker response, as well as the corresponding levels of MK1775 in the xenografts. Funded by NCI Contract No. HHSN261200800001E.

Citation Format: Deborah F. Wilsker, Allison M. Marrero, Melinda Hollingshead, Scott M. Lawrence, Alice Chen, Shivaani Kummar, Joseph M. Covey, Ralph E. Parchment, Robert J. Kinders, Joseph E. Tomaszewski, James H. Doroshow. Pre-clinical investigation of the wee1 inhibitor MK-1775 using pharmacodynamic and mechanistic markers in diverse cancer models in vivo. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4689. doi:10.1158/1538-7445.AM2014-4689
Abstract 4693: Inhibition of BET bromodomain targets PTEN positive endometrioid endometrial cancers
Authors: Qiu Haifeng , Jackson Amanda , Kilgore Joshua , Zhou Chunxiao , Bae-Jump Victoria Lin
Published: May 2015

Introduction: Endometrioid endometrial cancers (EEC) are commonly characterized by PTEN mutations and loss of function. JQ-1 is a selective small-molecule BET bromodomain inhibitor, and BET inhibition by JQ-1 leads to the downregulation of c-Myc transcription. Thus, JQ-1 functions as a novel c-Myc inhibitor and has been found to potently suppress tumor growth in many human cancers. Given the known interactions between c-Myc and the PI3K/Akt/mTOR pathway, we aim to assess the impact of JQ-1 on human EEC cell lines and further explore the mechanism underlying cellular resistance to JQ-1 treatment.

Methods: Six EEC cancer cell lines were used in this study. Cell proliferation was assessed by MTT assay and colony formation assay after exposure to JQ-1. Cell cycle progression was evaluated by flow cytometry. Apoptosis was assessed by Annexin V-FITC assay. Expression of c-Myc, PTEN, phosphorylated-Akt, phosphorylated-S6, cyclins and cyclin-dependent kinases (CDKs) was detected by Western blotting analysis. Up- or down-regulation of PTEN was achieved by stable transfection with pcDNA3.1-PTEN or shRNA-PTEN, respectively.

Results: JQ-1 significantly suppressed proliferation in the EEC cell lines that were PTEN positive (IC50=75 nM for HEC-1A, 550 nM for KLE and 1000 nM for ECC-1), but not in those that were PTEN negative (IC50>10000 nM for Ishikawa, AN3CA and RL 95-2). JQ-1 induced G1 cell cycle arrest in the PTEN positive HEC-1A and KLE cell lines, but had minimal effects on cell cycle progression in the PTEN negative Ishikawa and AN3CA cell lines. Western blotting analysis demonstrated that JQ-1 suppressed the expression of c-Myc, cyclins and CDKs. JQ-1 up-regulated PTEN expression and decreased phosphorylation of downstream targets of the PI3K/Akt/mTOR signaling pathway, including Akt and S6. Knock-down of PTEN led to cellular resistance to JQ-1 in the HEC-1A and KLE cells, while over-expression of PTEN sensitized Ishikawa and AN3CA cells to JQ-1 treatment.

Conclusions: We find that JQ-1 significantly suppressed cellular proliferation through G1 cell cycle arrest in the EEC cell lines with abundant PTEN expression. Knock-down of PTEN caused cellular resistance while over-expression of PTEN sensitized EEC cells to JQ-1. This evidence suggests that JQ-1 may be a promising targeted therapy for EEC patients with PTEN positive tumors.

Note: This abstract was not presented at the meeting.

Citation Format: Haifeng Qiu, Amanda Jackson, Joshua Kilgore, Chunxiao Zhou, Victoria Lin Bae-Jump. Inhibition of BET bromodomain targets PTEN positive endometrioid endometrial cancers. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4693. doi:10.1158/1538-7445.AM2014-4693
Abstract 4696: Identification of driver mutations and gene amplifications in resected asophageal adenocarcinoma: Impact on clinical care
Authors: Zhou Haiyu , Vargas Luis Tapias , Elliot Justin , Mathisen Douglas J. , Lanuti Michael
Published: May 2015

BACKGROUND:

Adenocarcinoma of the distal esophagus and esophagogastric junction (EGJ) have received much attention because of increasing frequency and poor outcome despite multimodality therapy. Driver mutations and gene amplifications in resected esophageal adenocarcinoma are not widely investigated. Opportunity exists to discover actionable mutations in resected esophageal adenocarcinoma.

METHODS:

We analyzed driver mutations harbored in coding regions of 25 selected genes (∼95 point mutations) using a multiplex PCR-based assay. EGFR, HER2, MET, and PI3K amplifications were tested using fluorescent in situ hybridization in patients who underwent esophagectomy for distal esophagus and EGJ adenocarcinoma from 2009 to 2013. Disease free survival (DFS) and overall survival (OS) were compared between patients with TP53, KRAS, PI3K, BRAF, APC mutant and wild-type tumors using Kaplan-Meier methods and Cox regression models. The impact of HER2 expression and outcomes were also analyzed.

RESULTS:

Of 54 distal esophagus and EGJ adenocarcinoma patients, 5 different driver mutations were observed in 39 patients, including TP53 (11/39, 28.2%), KRAS (6/39,15.4%), PI3K (2/11,5.1%), BRAF(1/39, 2.6%) and APC (2/39,5.1%). Double driver mutations (3/39, 7.9%) were shared in 3 patients with TP53 and KRAS (1/39, 2.6%), TP53 and PI3K (1/39, 2.6%), KRAS and PI3K (1/39, 2.6%), respectively. Amplifications in HER2 (10/45, 22.2%), MET (1/34, 2.9%) and EGFR (2/17, 11.8%) were observed where directed therapy was altered in 5 patients (1 patient received induction herceptin and 4 patients received adjuvant herceptin). In the targeted patients with HER2 amplification who received herceptin, mean progression-free survival was 18.8 (range 7-41 months) vs. 8.9 (1-28 months) [p=0.089] and OS was 35.7 (range 26-46 months) vs. 24.2 (range 20-29 months) [p=0.053], compared to herceptin naive patients. We did not identify any significant difference in DFS and OS (inclusive of all stages) between TP53, KRAS or any mutation compared to wild type patients.

DISCUSSION:

Oncogenic driver mutations are under-investigated in resected distal esophageal and EGJ adenocarcinoma. Different therapeutic opportunities may become available if routine tumor genotyping is employed in the treatment of esophageal adenocarcinoma. We recommend the routine use of driver mutation and gene amplification analysis for esophageal cancer.

Citation Format: Haiyu Zhou, Luis Tapias Vargas, Justin Elliot, Douglas J. Mathisen, Michael Lanuti. Identification of driver mutations and gene amplifications in resected asophageal adenocarcinoma: Impact on clinical care. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4696. doi:10.1158/1538-7445.AM2014-4696
Abstract 4697: Quantitation of BRAF V600E alleles predicts papillary thyroid cancer progression
Authors: Jung Chan Kwon , Kim Min Hee , Bae Ja Seong , Lim Dong Jun , Lee Hyoungnam , Jeon So Ra , Park Gyeong Sin
Published: May 2015

BACKGROUND: BRAF V600E is the most common genetic alteration in thyroid cancer. However, there is still controversy regarding its clinicopathological significance and clonal mutation frequency. To clarify these inconsistent results, we investigated the association between the allelic frequency of BRAF V600E and the clinicopathological features in classic papillary thyroid carcinoma (PTC).

DESIGN: Tumor tissues from 321 patients with classic PTC were microdissected under a dissecting microscope and analyzed for the presence or absence of the BRAF mutation and mutant allelic percentage using quantitative pyrosequencing.

RESULTS: BRAF V600E was found in 264 (82.2%) of 321 classic PTC. In the BRAF V600E-positive tumors, the mutant allelic frequency varied from 8% to 41% of total BRAF alleles (median, 20%). The frequency of BRAF V600E alleles directly correlated with tumor size (R-squared = 0.162, P<0.001) and number of metastatic lymph nodes (R-squared = 0.063, P<0.001). Out of 264 BRAF-positive PTCs, 136 (51%) had ≥20% of BRAF mutant alleles. Extrathyroidal extension and lymph node metastasis were more frequent in PTCs with high (≥20%) abundance of mutant alleles than in those with low abundance of mutant alleles (55% vs 37%, P=0.003 and 62% vs 46%, P=0.012, respectively). However, if the mutant allelic percentage was not considered, the presence of BRAF V600E was only associated with extrathyroidal extension, but not with tumor size and lymph node metastasis.

CONCLUSION: High abundance of BRAF V600E alleles predicts tumor progression in classic PTC. We suggest that the clonal, subclonal, or oligoclonal occurrence of BRAF V600E mutation may have different biological and clinicopathological significance in the PTC.

Citation Format: Chan Kwon Jung, Min Hee Kim, Ja Seong Bae, Dong Jun Lim, Hyoungnam Lee, So Ra Jeon, Gyeong Sin Park. Quantitation of BRAF V600E alleles predicts papillary thyroid cancer progression. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4697. doi:10.1158/1538-7445.AM2014-4697
Abstract 4703: Genome & exome analysis of early colon cancers reveals new targets
Authors: Majumdar Avijit , Chen Jian , Lin Sue Hwa , Katz Lior , Raju Gottumukkala Subba , Shetty Kirti , White Jon , Wu Xifeng , Rashid Asif , McMurray John S. , Shaw Kenna R. , Su Xiaoping , Weston Brian , Thirumurthi Selvi , He Aiwu Ruth , Mishra Lopa
Published: May 2015

Introduction: Advanced colorectal cancer (CRC) remains lethal and difficult to treat with over 140,000 new cases a year in the United States. One contributing factor that has emerged is the rapid development of cancers in small, sessile adenomas(polyps) (less than 1cm), which might be overlooked.

Results: Using whole genome and whole exome sequence (WGS) analyses of four pairs of colorectal adenoma (early CRC) tissue samples along with their normal counterparts, we observe dramatic genomic instability in two of the samples, suggesting that these two adenomas could be on the brink of overt CRC. We proceeded to analyze a further 20 samples through RNA sequence and/or immunohistochemical analysis. The two polyps are associated with hyper mutated profile of multiple genes including Carcinoembryonic antigen (CEA) and CEACAM6 in one of the samples. Analyses of the expression levels of CEA and TGF-β signaling pathway members in 20 early adenomas and normal colon tissues demonstrate a marked increase in CEA expression in 25% of adenoma samples linked to a concomitant loss of TGF-β signaling. We proceeded to mechanistic studies examining CEA and it role in inactivating the tumor suppressive arm of transforming growth factor-β (TGF-β) signaling, providing crucial new information for CEA in promoting aggressive CRC. We found that CEA interacts directly with the transforming growth factor-β (TGF-β) receptor 1 (TBR1) with inhibition of downstream TGF-β tumor suppressor signaling through its B3 domain. Moreover, we previously found that deletion of Smad3/4 adaptor β2SP results in loss of TGF-β signaling, with dramatic and spontaneous formation of adenomas as well as CRC. Ectopic expression of CEA also induces CRC metastases, potentially indirectly through activation of STAT3 and stimulating production of IL-6, IL-10 and others, in addition to inactivating TGF-β tumor suppressor signaling. Conclusions: Our data suggest that enhanced CEA levels and oncogenic mutations in the colon adenomas can be used as prognostic markers for early detection of aggressive CRC and the B3 domain of CEA can be targeted by monoclonal antibody/peptide inhibitors/small molecule inhibitors targeting CEA and CRC cell proliferation.

Citation Format: Avijit Majumdar, Jian Chen, Sue Hwa Lin, Lior Katz, Gottumukkala Subba Raju, Kirti Shetty, Jon White, Xifeng Wu, Asif Rashid, John S. McMurray, Kenna R. Shaw, Xiaoping Su, Brian Weston, Selvi Thirumurthi, Aiwu Ruth He, Lopa Mishra. Genome & exome analysis of early colon cancers reveals new targets. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4703. doi:10.1158/1538-7445.AM2014-4703
Abstract 4704: Prognostic significance of T gene SNP s2305089 in individuals with spinal column chordoma
Authors: Varga Peter , Fisher Charles , Rhines Lawrence , Gokaslan Ziya , Boriani Stefano , Lazary Áron , Wang Wei-Lien ‎ , Germscheid Niccole , Yip Stephen , Bettegowda Chetan
Published: May 2015

Chordoma is the most common primary spinal column tumor with a median survival of only 6-7 years, with extremely high recurrence rates. Radical surgical resection remains the mainstay of treatment for these tumors, with radiation often utilized to enhance local disease control. There are no approved chemotherapeutic agents for chordomas. Recently, Pillay and colleagues found that the single nucleotide polymorphism (SNP) rs2305089 in the T gene was strongly associated with chordoma development1. The variant A allele at rs2305089 results in a p.GLy177Asp alteration in brachyury, the protein encoded by the T gene. Brachyury has been implicated in the pathogenesis of chordomas and is highly expressed in most of these tumors. Given the rarity of this disease, with an incidence of 0.08 per 100,000 individuals, determining the clinical significance of this SNP has been challenging. In order to address whether the rs2305089 SNP was associated with altered prognosis in individuals with spinal column chordomas, we performed a retrospective analysis on 82 chordomas from 5 different high volume centers in Europe and North America. Each sample was linked to well curated clinical outcomes data that were collected after receiving institutional review board approval. The A variant at rs2305089 was present in 74 of 82 cases analyzed. In our cohort, there were no differences in gender, age, location, ethnicity, histologic subtype or time to local recurrence based on the presence of the A variant. However, overall survival was significantly worse (log-rank = 0.003) in individuals who lacked the A variant. To our knowledge, this is the first time the rs2305089 SNP has been implicated in the prognosis and survival of individuals with chordoma. This finding warrants further investigation and may have important implications on clinical decision-making.

1Nature Genetics. 2012 Nov;44(11):1185-7

Note: This abstract was not presented at the meeting.

Citation Format: Peter Varga, Charles Fisher, Lawrence Rhines, Ziya Gokaslan, Stefano Boriani, Áron Lazary, Wei-Lien ‎ Wang, Niccole Germscheid, Stephen Yip, Chetan Bettegowda. Prognostic significance of T gene SNP s2305089 in individuals with spinal column chordoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4704. doi:10.1158/1538-7445.AM2014-4704
Abstract 4705: Integrating germline cancer predisposition gene testing into routine clinical practice - The Mainstreaming Cancer Genetics programme
Authors: Rahman Nazneen ,   The Mainstreaming Cancer Genetics Consortium
Published: May 2015

Over 100 cancer predisposition genes with clinical utility are recognised. Testing of these genes is increasingly in demand and is one of the major activities of Clinical Genetics. However, in most countries, testing is very restricted with respect to the number of genes and the number of people tested. Furthermore, the value of genetic testing in individuals with cancer is underappreciated; it provides important information with respect to the cause and optimal treatment of the current cancer, and the risk and optimal management of future cancer. Moreover, testing cancer patients followed by cascade testing of mutation carriers is an effective and efficient way of identifying unaffected mutation carriers in whom screening and risk-reducing strategies can be deployed. The Mainstreaming Cancer Genetics (MCG) Programme is a UK national cross-disciplinary initiative to develop the NGS assays, analytical and interpretive pipelines, clinical infrastructure, training, ethical and evaluation processes required for routine genetic testing to be integrated into cancer patient care.

In collaboration with Illumina we have developed a NGS panel targeting 97 cancer predisposition genes with >1500 probes (called the TruSight cancer panel) and have completed detailed evaluation of performance, sensitivity and specificity under different levels of multiplexing, coverage and throughput. We have developed bespoke analytical pipelines developed for high-throughput clinical diagnostic data analysis (called GAMA) and clinical interpretation (called CIGMA). We are implementing a new mixed-model of cancer gene testing whereby consent for medical testing (i.e. in cancer patients) is undertaken by trained oncologists, with only the mutation carriers seen by geneticists. Predictive testing (i.e. in unaffected individuals) continues to be undertaken in genetics. We have developed protocols and e-learning modules to deliver the required training for oncologists to consent for testing. The system is faster and cheaper, whilst retaining flexibility and robustness to provide increased input where required. We are completing a 6 month pilot that has included >100 patients and we are acquiring patient and clinician feedback, the results of which will be presented. www.mcgprogramme.com

Citation Format: Nazneen Rahman, The Mainstreaming Cancer Genetics Consortium. Integrating germline cancer predisposition gene testing into routine clinical practice - The Mainstreaming Cancer Genetics programme. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4705. doi:10.1158/1538-7445.AM2014-4705
Abstract 471: Transcriptional coactivator CBP regulates hTERT expression through the cooperation with Sp1/AP-2β and predicts poor prognosis in human lung cancers
Authors: Guo Wei
Published: May 2015

Human telomerase reverse transcriptase (hTERT) is linked to tumorigenesis and development. However, how hTERT is upregulated in tumor cells remains poorly understood. In this study, we identified CBP as a transcriptional coactivator that specifically binds to hTERT promoter through co-anchoring with transcriptional factors Sp1 and AP-2β and promotes hTERT expression in human lung cancer cells. Streptavidin-agarose pulldown and chromatin immunoprecipitation assays revealed the tumor-specific binding of CBP, Sp1 and AP-2β on the hTERT promoter. The colocalization and interaction between CBP and Sp1 or AP-2β in lung cancer cells were detected. High expression levels of CBP and hTERT in lung cancer cells and tissues were also detected compared to normal lung cells and adjacent normal lung tissues. Tissue array immunohistochemistry of lung adenocarcinomas showed a strong postive correlation between CBP and hTERT expression, and demonstrated the patients with high CBP and hTERT expression had a significantly shorter overall survival than those with low CBP and hTERT expression. Ectopic expression of CBP activated hTERT promoter-driven luciferase gene and endogenous hTERT gene expression in lung cancer cells. Inhibition of CBP expression by CBP-specific siRNA or chemical inhibitor attenuated hTERT promoter-driven luciferase expression and the endogenous hTERT activity, and abrogated the binding of Sp1 and AP-2β on hTERT promoter region. Moreover, CBP knockdown by siRNA significantly inhibited lung cancer cell proliferation, whereas CBP overexpression promoted cell proliferation. These results therefore indicate overexpression of CBP contributes to hTERT upregulation through its cooperation with transactivators Sp1 and AP-2β and predicts poor prognosis in human lung cancers.

Note: This abstract was not presented at the meeting.

Citation Format: Wei Guo. Transcriptional coactivator CBP regulates hTERT expression through the cooperation with Sp1/AP-2β and predicts poor prognosis in human lung cancers. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 471. doi:10.1158/1538-7445.AM2014-471
Abstract 4724: Adverse prognostic significance of TNFAIP8 protein expression in non small cell lung cancer
Authors: Kallakury Bhaskar V. , Kasid Usha N. , Voronel Olga , Sheehan Christopher B. , Sheehan Christine E. , Ross Jeffrey S.
Published: May 2015

Background: Tumor necrosis factor alpha induced protein 8 (TNFAIP8) is a transcription factor NF-kB inducible antiapoptotic and oncogenic molecule. Positive correlation between TNFAIP8 expression and cancer progression/poor prognosis has been described in a few human solid cancers including pancreatic and esophageal primaries. We were the first to publish the correlation of TNFAIP8 expression with high tumor grade and its independent predictor status for disease recurrence in prostatic adenocarcinomas. In this study, we evaluated the significance of this protein in non small cell lung cancer (NSCLC).

Design: A total of 144 formalin-fixed, paraffin-embedded NSCLCs were analyzed (122 primary tumor resections and 22 tissue microarray samples); including 47 squamous cell carcinomas (SCC), 59 adenocarcinomas (AC), and 38 bronchioloalveolar carcinomas (BAC), were immunostained by a manual method using a custom made anti-TNFAIP8 antibody as described earlier (Zhang et al., Int J Cancer,133: 31-42, 2013). The staining pattern was semiquantitatively assessed based on staining intensity and distribution and the results were correlated with morphologic and prognostic variables.

Results: Immunoreactivity for TNFAIP8 was predominantly cytoplasmic. TNFAIP8 protein overexpression was noted in 84/144 (58%) tumors and correlated with tumor type [45/59 (76)% AC vs 29/47 (62%) SCC vs 10/38 (26%) BAC, p<0.0001], overall with advanced tumor stage [24/31 (77%) advanced vs 60/113 (53%) early, p=0.015] and lymph node positive status [31/41 (76%) LN+ vs 53/103 (52%) LN-, p=0.008]. Within the AC subgroup, there was a trend toward correlation with advanced tumor stage [13/14 (93%) advanced vs 32/45 (71%) early, p=0.095] and lymph node positive status [18/20 (90%) LN+ vs 27/39 (69%) LN-, p=0.076]. On multivariate analysis, LN+ status independently predicted shortened survival.

Conclusion: TNFAIP8 immunohistochemical expression portends poor prognosis in NSCLC and may constitute a novel therapeutic target in the management of aggressive NSCLC, warranting further study.

Citation Format: Bhaskar VS Kallakury, Usha N. Kasid, Olga Voronel, Christopher B. Sheehan, Christine E. Sheehan, Jeffrey S. Ross. Adverse prognostic significance of TNFAIP8 protein expression in non small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4724. doi:10.1158/1538-7445.AM2014-4724
Abstract 4727: Prognostic roles of DDB2 and XPC mRNA expressions in treatment responses and overall survivals of head and neck cancer patients
Authors: Lin Chang-Shen , Lee Ka-Wo , Huang Jau-Ling , Wang Yu-Chu , Tsai Yi-Shan
Published: May 2015

Background:

DNA repair genes function in the maintenance of genome integrity and prevent tumorigenesis. On another hand, DNA repair genes modulate cancer cell responses to DNA-damaging therapeutics. We hypothesize that altered expression of DNA repair genes are correlated with therapeutic outcomes of head and neck cancer patients.

Patients and methods:

Messenger RNA (mRNA) expressions of DDB2 and XPC were examined in specimens of 95 head and neck cancer patients using real-time quantitative RT-PCR. Patients were dichotomized by ROC curve analysis based on their DDB2 and XPC mRNA levels. Overall survival (OS) rates were measured using Kaplan-Meier estimates and the log-rank tests. The adjusted hazard rate ratios (HRRs) were computed by multivariate Cox regressions.

Results:

The levels of DDB2 and XPC mRNA expression in tumor tissues were reduced (tumor/normal < 0.9) in 84 and 80 cases, respectively, when compared with those in adjacent normal parts. Lower DDB2 (tumor/normal < 0.24) and XPC (tumor/normal < 0.36) mRNA expressions were correlated with larger tumor, positive lymph node invasion, advanced stage, and poor 5-yr OS rates (26.7% vs. 54.3%, and 28.1% vs. 53.0%, respectively). Multivariate Cox model showed that, in addition to T and N stages, lower XPC mRNA level (< 0.36) was an independent risk factor for patient's survival (HRR = 2.68, 95% CI 1.12-6.40). Concurrent chemotherapy and radiotherapy (CCRT) did not improve OS of patients with lower XPC mRNA (< 0.36) and showed even worse outcomes within 5 years for patients with lower DDB2 expression (< 0.24). However, patients with higher DDB2 (> 0.24) and XPC (> 0.36) mRNA levels had improved 5-yr OS rates if they received CCRT (64.4% vs. 41.8% and 73.3% vs. 39.2%).

Conclusion:

mRNA expressions of XPC and DDB2 might be useful biomarkers for prognosis and efficacy of CCRT medication in head and neck cancer patients.

Citation Format: Chang-Shen Lin, Ka-Wo Lee, Jau-Ling Huang, Yu-Chu Wang, Yi-Shan Tsai. Prognostic roles of DDB2 and XPC mRNA expressions in treatment responses and overall survivals of head and neck cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4727. doi:10.1158/1538-7445.AM2014-4727
Abstract 4728: FGFR1 gene amplification in small cell lung cancer
Authors: Park Ji Soo , Lee Jae-Seok , Shim Hyo Sup , Kim Hye Ryun , Lim Sun Min , Kim Joo Hang , Cho Byoung Chul
Published: May 2015

Background: Fibroblast growth factor receptor 1 (FGFR1) received special attention from many clinicians according to finding that FGFR1 played a critical role in many human cancers. Some clinical trials using FGFR1 targeting agents are in progress especially in breast cancer and squamous non-small cell lung cancer. Recently, FGFR1 amplifications were also found in about 5.6-6% of small cell lung cancer (SCLC) by gene copy number analysis and FISH.

Methods: Tumor tissues from 113 patients diagnosed with SCLC from October 2009 to February 2013 in YUHS, Korea were collected. FGFR1 FISH assay was performed on the tissue microarray using FISH probe that hybridized to the band 8p12-8p11.23 (FISH probes were provided by Abbott Molecular, Abbott Park, IL).

Results: Thirty-two patients with limited-stage disease (LD) and eighty-one patients with extensive-stage disease (ED) were enrolled. Fourteen patients were female. Among 113 tumor samples, 5 samples (4.4%) showed FGFR1 amplification. All 5 samples were obtained from the patients with ED. The patients with FGFR1 amplification more frequently had pleural metastasis at the time of diagnosis (80% vs. 26%, P=0.022). In ED, the patients with FGFR1 amplification represented shorter disease free survival from first line chemotherapy (Hazard ratio [HR], 4.469; 95% confidence interval [CI], 1.600-12.483; P=0.004) and second line chemotherapy (HR, 6.533; 95% CI, 1.700-25.108; P=0.006) than the patients without FGFR1 amplification in multivariate analysis. Median overall survival time was 8.2 months in the patients with FGFR1 amplification and 10.2 months in the patients without FGFR1 amplifications (P=0.296).

Conclusion: FGFR1 amplification is a poor potential predictive biomarker of standard chemotherapy, necessitating further research in a large cohort of SCLC. The validity of targeting FGFR1 in SCLC should be confirmed in a clinical trial.

Citation Format: Ji Soo Park, Jae-Seok Lee, Hyo Sup Shim, Hye Ryun Kim, Sun Min Lim, Joo Hang Kim, Byoung Chul Cho. FGFR1 gene amplification in small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4728. doi:10.1158/1538-7445.AM2014-4728
Abstract 4735: Stem cells like phenotype of inflammatory breast cancer and locally advanced breast cancer: increased expression of sox2 and oct3/4 contributes to poor response to treatment and reduced survival rates
Authors: Zekri Abdel-Rahman N. , Bahnassy Abeer , El-Bastawisy Ahmed E. , Nassar Hanan R. , Elhady Neveen M. , Effat John
Published: May 2015

Background: Inflammatory breast cancer (IBC) is an aggressive form of locally advanced breast cancer (LABC) with poor response to treatment or metastasis in one-third of patients at presentation. The aggressive behavior of IBC was attributed to the cancer stem cell (CSC) component in these tumors. We assessed the correlation between CSC markers in IBC, LABC and the aggressive behavior of these tumors compared to invasive duct carcinoma (IDC-NOS).

Methods: CD44, CD90, CD133, nestin, SOX2 and OCT4 RNA and proteins expression were assessed in 30 cases of IDC-NOS, IBC and LABC (each) using quantitative real time PCR (qPCR) and immunohistochemistry (IHC). Markers expression was correlated to patients' characteristics, response to treatment, 4 years- overall and progression free survival (OS& PFS) .

Results: The expression levels in IBC and LABC was as follows: (CD44; 70%& 26.3%, p=0.003), (CD90; 49.2%& 28.1%, p=0.06), (CD133; 66.7%& 26.3%, p=0.006), (OCT4; 70%& 26.6%, p=0.004), (SOX2: 63.3%& 30%, p=0.02) and (Nestin; 53.3%& 31%, p=0.08). The difference in the expression levels of all studied markers between IDC-NOS and the other two groups was statistically significant (p.value: <0.05- <0.001). A significant relation was reported between the expression levels of SOX2, OCT-4 and the clinical response in IBC and LABC (p<0.01). PFS significantly correlated with SOX2 and OCT4 expression (p=0.045) in IBC, whereas OS correlated with OCT4 in IBC and LABC (0.034& 0.04)

Conclusions: The aggressive phenotype of IBC and LABC could be attributed, at least partially, to the presence of CSC populations in these tumors. Therefore, CSC markers represent Interesting candidates for targeted therapy for this resistant type of breast tumors.

Citation Format: Abdel-Rahman N. Zekri, Abeer Bahnassy, Ahmed E. El-Bastawisy, Hanan R. Nassar, Neveen M. Elhady, John Effat. Stem cells like phenotype of inflammatory breast cancer and locally advanced breast cancer: increased expression of sox2 and oct3/4 contributes to poor response to treatment and reduced survival rates. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4735. doi:10.1158/1538-7445.AM2014-4735
Abstract 4737: Role of hypoxia-inducible factor-1α, carbonic anhydrase-IX and glucose transporter-1 in cervical cancer
Authors: Iwasaki Keita , Yabushita Hiromitsu , Ueno Taiki , Wakatsuki Akihiko
Published: May 2015

Objectives: Hypoxia can drive the tumor toward a more aggressive malignant phenotype through genomic and proteomic changes. Hypoxia-inducible factor-1α (HIF-1α) transactivates genes whose proteins allow metabolic adaptation to hypoxia, including carbonic anhydrase-IX (CA-IX) and glucose transporter-1 (Glut-1). This study aimed to determine whether the HIF-1α, CA-IX and Glut-1 were associated with clinico-pathological characteristics and progression-free survival in patients with cervical carcinoma.

Methods: Under the written informed consent, tumor tissue samples were obtained from 54 patients with cervical carcinoma performed radical hysterectomy. The expressions of HIF-1α, CA-IX and Glut-1 were detected by immunohistochemical staining using paraffin-embedded sections.

Results: CA-IX and Glut-1 expressions were found in cell membrane and cytoplasm, and HIF-1α expression was observed in cell nuclear. In 54 cases, the positive expression was observed in 28 cases for HIF-1α, 35 for CA-IX and 40 for Glut-1. The HIF-1α expression was related with clinical stage and histology, CA-IX expression was related with clinical stage, tumor size, lymph-node metastasis and lymph-vascular space involvement, and the Glut-1 expression was related with clinical stage and lymph-vascular space involvement. The multivariate regression analysis revealed that the CA-IX expression and lymph-vascular space involvement were the independent variable associated with lymph-node metastasis in patients with cervical cancer. Progression-free survival was shorter in patient with positive CA-IX expression compared to negative CA-IX expression.

Conclusion: In cervical cancer patients, CA-IX expression is possible to be a risk factor for lymph-node metastasis and disease recurrence, it was suggested that pH regulation induced by CA-IX under hypoxic condition may be associated with lymph node metastasis.

Citation Format: Keita Iwasaki, Hiromitsu Yabushita, Taiki Ueno, Akihiko Wakatsuki. Role of hypoxia-inducible factor-1α, carbonic anhydrase-IX and glucose transporter-1 in cervical cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4737. doi:10.1158/1538-7445.AM2014-4737