Abstract Background: Triple negative breast cancer (TNBC) is a heterogeneous disease with several molecular subtypes: basal-like1 (BL1), basal-like 2 (BL-2), mesenchymal(M), mesenchymal-stem-like(MSL), immune-modulatory(IM) and unclassified (UNC) Molecular evolution of TNBC through chemotherapy selection pressure is well recognized but poorly understood. This study was carried out to perform paired genomic analysis of TNBC comparing primary breast cancer with recurrent/refractory disease. Here we report the result of the first10 paired tissue analysis. Methods: 49 paired specimens were identified through an IRB-approved protocol via COH biorepository search (2002- 2015). miRNA and mRNA profiling of 22 samples were performed. The miRNA libraries were prepared and sequenced on Hiseq2500. Sequences were aligned to hg19 genome and miRNA expression levels were counted by in house built R scripts. Go and pathway annotation was performed using DAVID online tool. Affymetrix human Genechip 2.0st was used for mRNA expression profiling. Raw data were normalized and processed using Expression Console, and linear regression was performed using Limma to identify the differentially expressed genes between primary and recurrent/refractory TNBCs. Result: Through mRNA profiling, we identified several unique gene expression patterns comparing the paired TNBC. Significant mRNA expression alterations were observed in: cell cycle, DNA repair and adhesion. Using Vanderbilt TNBC sub-classification tool, we have identified “phenotype shift” between primary and recurrent TNBCs. Of the 8 paired specimen analyzed, 3 paired tissue remain in the same subclass (2 in IM, 1 in M). Phenotype shift observed in: 1 from BL1 to BL2, 1 from BL2 to BL1, 1 UNC to IM; 1 MSL to UNC; 1 from M to UNC. 15 up regulated and 13 down regulated miRNAs were identified. Most significantly differentially expressed miRNA (with more than 4 fold expression changes, P-value < 0.001) included: miR-206, miR-203, miR-144, miR-16-2, miR-15b, and miR-20b (un-regulated) and miR-10b, miR-125b and let-7c(down-regulated). These miRNA genes are involved in regulation of hormonal receptor signaling, cell cycle, proliferation and metastases. Statistically significant differentially expressed miRNAs identified from our TNBC patient cohort will be further validated using RT-PCR. Conclusion: A number of mRNA gene pathways and miRNAs showed differential expression between paired recurrent and primary TNBC tumor specimen. The underlying biology driven the phenotype shift is being studied. Further analysis to include a total of 49 paired TNBCs is currently underway. Contact information: Yuan Yuan MD PhD, Email: yuyuan@coh.org. Citation Format: Yuan Y, Li A, Yost S, Yuan Y-C, Chu P, Warden C, Wang J, Liu Z, Liang Y, Goldstein L, Wu X. Genomic analysis of molecular discordance of paired primary and recurrent triple negative breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-05-26.
9054 Background: The prognosis for patients with recurrent Ewing's sarcoma (ES) and primitive neuroectodermal tumors (PNET) remains poor. Novel therapies are urgently needed. We previously demonstrated that EFT cell lines express the c-Kit and PDGFR-α tyrosine kinase receptors (TKRs). Further, we noted that imatinib mesylate blocks the functional activation of these TKRs, and inhibits EFT clonogenicity (Proc AACR 43: 336a, 2002). Additionally, we reported that these TKRs are detectable by immunohistochemistry (IHC) in a majority of primary EFT and DSRCT (Proc ASCO 22: 826a). Based upon these findings, we treated a patient with recurrent EFT that expressed both c-Kit and PDGFR-α with imatinib mesylate. We report our experience. Methods: The index patient was enrolled in an IRB-approved clinical trial. She satisfied standard physiologic and performance criteria. The starting dose of imatinib mesylate was 400 mg BID. Standard dosage reductions were followed for hematologic and non-hematologic toxicities. Results: The index patient is a 29 y/o W/F who presented in August, 2002 with back and abdominal pain. CT scan in October, 2002 revealed a large retroperitoneal mass, extensive retroperitoneal adenopathy, and ascites; a biopsy revealed PNET. Staging evaluation revealed bone and bone marrow involvement. Chemotherapy with vincristine, doxorubicin and cyclophosphamide, altenating with ifosfamide and etoposide was initiated. Concomitant radiation was offered during the sixth and seventh cycles. Progression was documented after 11 cycles of chemotherapy. She required long-acting morphine (160 mg every 8 hours) for pain. IHC of the original tumor revealed 3+/3 IHC positivity for both c-Kit and PDGFR-α. She was enrolled on the present trial on April 28, 2003. Within 2 weeks she discontinued morphine. Partial remission (RECIST criteria) was documented on October 21, 2003. She presently continues in partial response. Conclusions: This case report demonstrates that imatinib mesylate was an effective alternative therapy for this patient with recurrent EFT whose tumor expressed the c-Kit and PDGFR-α TKRs. Imatinib mesylate should be further studied in this area (Sponsored by Novartis). Author Disclosure Employment or Leadership Consultant or Advisory Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Novartis Novartis
9569 Background: We set out to establish prognostic and predictive indicators of outcome in patients (pts) with high-risk primary breast cancer (HRBC), by evaluating pathologic and molecular features of the primary tumors (PT). Methods: RNA was extracted from paraffin-embedded PT for RT-PCR testing for a panel of genes, including EGFR, HER-2, GST-Pi, ERCC1, p21, and Beta tubulin-3. H&E and immunohistochemical stain (IHC) were examined to assess expression of 12 proteins, including p16, p21, p27, HER-2, p53, ER/PR, CD1a, and Ki-67. The analysis was carried out in 240 (52 % of all eligible) pts who were treated on IRB-approved platinum, doxorubicin (Dox) and/or paclitaxel-containing dose-intense chemotherapy (DICT) protocols between 1989 and 2003, at the COHNMC. Results: Pts were treated for stage II ( ≥ 10 lymph nodes with metastasis [LNM]; 41%), IIIA (39%), and IIIB inflammatory BC (20%). The median age was 46 years (26–62), the median no. of LNM was 12 (0–42). Twenty percent of pts received neoadjuvant Dox CT with or withour a taxane. All pts received adjuvant CT with Dox with or without a taxane, followed by DICT. At a median follow-up of 86 months RFS and overall survival (OS) are 50% (95% CI; 43–57%) and 62% (56–69%). In univariate analysis RFS was shorter in pts whose PT were characterized by high-grade, ER/PR receptor negativity, greater expression of p16 by IHC, and low expression of p21 by IHC and by RT-PCR, and increased EGFR expression; OS was similarly effected by the same parameters and by lack of p27 and overexpression of Ki-67 by IHC (p< 0.05). In multivariate Cox stepwise analysis, after adjusting for age and stage, p21 overexpression and ER/PR receptor positivity were associated with improved RFS and OS. The predictive value of expression of other drug-resistance genes for specific DICT-associated survival is under evaluation. Conclusion: A prognostic model combining RT-PCR and IHC detection of markers of cell regulation, proliferation, and resistance may help to define target-specifc therapies in patients with HRBC. Author Disclosure Employment or Leadership Consultant or Advisory Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Response Genetics Response Genetics Response Genetics; Bristol-Myers Squibb; Amgen
The normal function of the retinal pigment epithelium (RPE) is dependent on the maintenance of tight adhesions between cells. In order to identify cell surface molecules which may be important for maintaining the integrity of the RPE, we have undertaken a combined functional, biochemical, and immunohistochemical analysis of cell surface proteins of the RPE. These studies have led to the identification of a 100-kD cell surface protein whose presence correlates with the maintenance of calcium-dependent adhesions between RPE cells. In intact RPE tissue the protein is concentrated at the junctions between RPE cells. The properties of the protein suggest that it may be a member of the cadherin family of calcium-dependent cell adhesion proteins.
Abstract Background: Triple negative breast cancer (TNBC) is a heterogeneous disease with several molecular subtypes: basal-like1 (BL-1), basal-like 2 (BL-2), mesenchymal (M), and luminal androgen receptor (LAR). Molecular evolution of TNBC through chemotherapy selection pressure is well recognized but poorly understood. In addition, approximately 20% of TNBCs respond to PD-1 or PD-L1 inhibitors. It has been observed that heavily pre-treated patients may not respond well to immunotherapy. This study was carried out to perform immune profiling of paired primary and recurrent TNBC. Here we report the result of the first 10 paired tissue pilot analysis. Methods: Twenty specimens were identified through an IRB-approved protocol via the City of Hope Biospecimen Repository (2002-2015). Two brain and one bone metastasis specimens were not included due to technical difficulty. Formalin-fixed paraffin embedded (FFPE) sample blocks were cut into 5-mm thick slides and labeled with the following antibodies: CD4, CD8, CD3, FOXP3, CD20, CD33, Pan-CK, and PD-1 using the multiplex IHC opal method. Image acquisition and cell counting were carried out using PerkinElmer Vectra automated quantitative pathology imaging system and inForm software analysis (PerkinElmer, Waltham, MA). mRNA expression profiling was performed using Affymetrix Human Genechip 2.0. Raw data were normalized and processed using Expression Console. Using Vanderbilt TNBC sub-classification tool, we have sub-classified the 20 primary and recurrent TNBC specimens. Tumor mutation burden (TMB) was generated through FoundationOne® platform. Result: A total of 17 samples were analyzed (M, 5; LAR, 3; BL-1, 4; BL-2, 5). M-subtype had a significantly lower tumor-infiltrating CD3+ T cells (p=0.005), CD8+ T cells (p=0.024), CD4+ T cells (p=0.065) and CD4+FOXP3+ Treg cells (p=0.054), irrespective of the site of metastasis. CD20+ B cells were particularly enriched in BL-1 subtype (p=0.0013, 23.5% of 17 samples). Of 17 samples, 8 had TMB. Seven had low TMB (<10 mut/Mb) and one had intermediate TMB (11 mut/Mb, LAR subtype). The tumor with intermediate TMB had the highest quantity of tumor-infiltrating CD3+ T cells, CD8+ T cells, CD8+PD1+ T cells, and CD4+FOXP3+PD1+ Treg cells compared to the 7 tumors with low TMB. Compared with recurrent tumors, primary tumors had a significantly higher percentage of tumor-infiltrating T cells (TIL). To validate multiplexed IHC results, these samples were evaluated by a licensed pathologist at City of Hope using the International TILs Working Group 2014 guidelines, and there was a good correlation between percent of TILs and CD3+ T cells by IHC approach. Conclusion: To our knowledge, this is the first study linking tumor immune cell profiles with the TNBC 4 subtypes. Distinctive immune cell patterns were observed among 4 TNBC subtypes. M subtype had significantly lower TILs, which may indicate poor response to checkpoint inhibitors. Further analysis of a total of 50 paired TNBCs is currently underway. Contact information: Yuan Yuan MD PhD, Email: yuyuan@coh.org Citation Format: He T-F, Yost S, Schmolze D, Wang R, Rosario A, Tu T, Chu P, Lee P, Yuan Y. Immune profiling of paired primary and recurrent triple negative breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P3-05-02.
Results 59-year-old gentleman was referred to clinic for weight loss of 10kg over 3months. He had appendicectomy and partial colectomy in mainland China 20years ago for intestinal obstruction, partial gastrectomy for perforated peptic ulcer 10years ago. Detailed operative record was not traceable.For years he has had on and off diarrhea, significant weight loss over the recent 3months. Blood tests showed hypokalemia, low B12, macrocytic anemia, hypoalbuminemia. Upper endoscopy showed Billroth II gastrectomy otherwise unremarkable. Barium meal and follow-through showed partial gastrectomy with normal transit time from stomach to colon. Colonoscopy showed poor bowel preparation with sticky stool obscuring mucosal surface; two small bowel-colon anastomoses at 50cm from anal verge. Unremarkable PET-CT and albumin scintiscan. He suffered an episode of severe hypoalbuminemia, hypokalemia, pulmonary edema complicated by septicemia, DIC necessitating intensive care.
Cytokeratin 14 expression in epithelial neoplasms: a survey of 435 cases with emphasis on its value in differentiating squamous cell carcinomas from other epithelial tumours Aims : The tissue distribution of cytokeratin 14 (CK14) in epithelial neoplasms is not well defined. We have evaluated 435 cases of epithelial neoplasm of various origins with cytokeratin 14 monoclonal antibody with special attention to possible use in differential diagnosis. Methods and results : Immunohistochemistry (ABC–HRP method) was performed for detection of CK14. We found that the expression of cytokeratin 14 was generally restricted to: (i) the majority of cases of squamous cell carcinoma regardless of origin (67/74) and degree of differentiation; (ii) neoplasms with focal squamous differentiation, including endometrial, and ovarian adenocarcinoma, malignant mesothelioma and transitional cell carcinoma; (iii) thymoma (8/8); (iv) myoepithelial components of salivary gland pleomorphic adenoma (3/4); and (v) oncocytic neoplasms, including thyroid Hurthle cell adenoma (1/1) and salivary gland Warthin’s tumour (2/2). Conclusion : CK14 protein is a useful marker in differential diagnosis of squamous cell carcinomas.
This book grew out of the authors' conversations over many years about how to improve family dispute resolution and the family justice system.We came to this work with different perspectives and experiences.We both have a foot firmly in the theory and practice of family law, but Jane's many years in clinical education and Jana's strong focus on scholarship brought different strengths and orientations to the work.This resulted in a collaboration that was, at times, challenging but always rewarding; we believe it made the book more balanced, thoughtful, and grounded in the real world.Both authors' views were enriched and changed by the process of writing this book.
