Prognostic Factor For Metastasis-free Survival Research Articles

Prognostic fine needle aspiration biopsy of uveal melanoma: Molecular and genetic factors of metastasis risk

Introduction. Molecular genetic testing is actively used for prognostication in patients with uveal melanoma (UM). Tissue for genetic analysis may be obtained either by surgical excision or through fine-needle aspiration biopsy (FNAB). Performing genetic testing and FNAB in each institution can differ in surgical techniques and laboratory methodologies.Aim. To present our own experience of performing FNAB-based molecular genetic testing for prognostication in patients with uveal melanoma.Materials and methods. Prognostic FNAB (n = 151) were combined with brachytherapy or stereotactic surgery. Genetic testing was performed by methods based on polymerase chain reaction (GNAQ, GNA11, EIF1AX and SF3B1 mutations) and fluorescence in situ hybridization (copy numbers of PPARG and MYC genes); cytology of FNAB material was also assessed.Results. Fine-needle aspiration biopsy material was informative in 91 % of cases. At the median follow-up of 36 months, 12 cases of distant metastases were detected. Occurrence of the assessed mutations and copy numbers were related to other representative studies. PPARG deletion was shown to be a significant prognostic factor for metastasis-free survival (p <0,01), which was demonstrated for the first time; EIF1AX and SF3B1 mutations, MYC amplification and cytological class were not shown to be significantly associated with survival in our study.Conclusion. FNAB-based molecular genetic testing for prognostication in patients with uveal melanoma was shown to be a reliable and highly informative approach. Some of the prognostic factors need to be evaluated further with longer follow-up.

CD105 expression in cancer-associated fibroblasts: a biomarker for bone metastasis in early invasive ductal breast cancer patients.

Introduction: Bone metastasis is one of the causes that mainly decrease survival in patients with advanced breast cancer. Therefore, it is essential to find prognostic markers for the occurrence of this type of metastasis during the early stage of the disease. Currently, cancer-associated fibroblasts, which represent 80% of the fibroblasts present in the tumor microenvironment, are an interesting target for studying new biomarkers and developing alternative therapies. This study evaluated the prognostic significance of the CD105 expression in cancer-associated fibroblasts in early breast cancer patients. Methods: Immunohistochemistry was used to assess CD105 expression in invasive ductal breast carcinomas (n = 342), analyzing its association with clinical and pathological characteristics. Results: High CD105 expression in cancer-associated fibroblasts was associated with an increased risk of metastatic occurrence (p = 0.0003), particularly bone metastasis (p = 0.0005). Furthermore, high CD105 expression was associated with shorter metastasis-free survival, bone metastasis-free survival, and overall survival (p = 0.0002, 0.0006, and 0.0002, respectively). CD105 expression also constituted an independent prognostic factor for metastasis-free survival, bone metastasis-free survival, and overall survival (p = 0.0003, 0.0006, and 0.0001, respectively). Discussion: The high CD105 expression in cancer-associated fibroblasts is an independent prognostic marker for bone metastasis in early breast cancer patients. Therefore, the evaluation of CD105(+) CAFs could be crucial to stratify BCPs based on their individual risk profile for the development of BM, enhancing treatment strategies and outcomes.

Abstract 2205: KRTAP 2-3 is a novel potential biomarker of cells in the polyaneuploid cancer cell (PACC) state to predict cancer recurrence

Abstract Prostate cancer (PCa) is the second leading cause of death in American men. PCa death is primarily attributed to therapeutic resistance. Our group and others have recently discovered a stress-induced adaptive cell state that plays a key role in therapy resistance: the polyaneuploid cancer cell (PACC) state. Cells in the polyaneuploid cancer cell (PACC) state are characterized by large cell size and polyploidization of the baseline aneuploid genome. Cells in the PACC state (e.g., pleomorphic giant cells, polyploid giant cancer cells, osteoclast like cells) have been reported in patient tumor specimens of multiple cancer types. We recently reported that in primary tumor specimens of PCa patients undergoing radical prostatectomy with curative intent, the number of cells in the PACC state is a statistically significant negative prognostic factor for metastasis-free survival (data adjusted for CAPRA-S). These data highlight the promise of the PACC state as a predictive biomarker for recurrence. In our previous work, cells in the PACC state were identified and quantified through EpCAM immunohistochemistry (IHC) to visualize cell membranes, manually including cells with &gt3x increased nuclear area compared to adjacent non-polyploid cancer cells. However, this method is labor intensive and highly subjective. There are currently no biomarkers to distinguish cells in the PACC state. To identify candidate PACC state biomarkers, we performed RNA sequencing to identify differentially expressed genes. Across two independent RNA sequencing datasets of PC3 cells induced to enter the PACC state with IC50 cisplatin or IC90 docetaxel, the most differentially expressed gene was KRTAP2-3 (Log2FC=4.3 and 5.51, respectively). The Keratin Associated Protein 2-3 (KRTAP2-3) is encoded by one of the homologous KRTAP genes classified by high sulfur content and is primarily expressed in the cortex of the hair follicle. It has not been previously implicated in PCa or in chemotherapy resistance. Due to its differential RNA expression in the PACC state and its low expression in most normal tissues, we hypothesize that KRTAP2-3 can be used as a biomarker to distinguish cells in the PACC state in patient tumor samples. To test this directly, we optimized RNA in situ-hybridization (RNAish) for KRTAP2-3. In initial experiments utilizing PC3 cell line plugs, we found that the KRTAP2-3 RNAish signal successfully distinguished cells in the PACC state (50.6 average punctae/cell) from non-polyploid cells (0.19 average punctae/cell) reflecting 96% percent accuracy using sensitivity and specificity metrics. We have further confirmed positive KRTAP2-3 signal on PC3 cells induced to access the PACC state in response to cisplatin, docetaxel, and etoposide treatment. Future work will evaluate the utility of the KRTAP2-3 gene as a prognostic biomarker that marks cells in the PACC state within PCa patient tissue microarrays. Citation Format: Sabrina A. Mendez, Reema Sharma, Kenneth J. Pienta, Sarah R. Amend. KRTAP 2-3 is a novel potential biomarker of cells in the polyaneuploid cancer cell (PACC) state to predict cancer recurrence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2205.

Predictors of Recurrence After Metastasis-directed Therapy in Oligorecurrent Prostate Cancer Following Radical Prostatectomy

BackgroundMetastasis-directed therapy (MDT) is performed to delay systemic treatments for oligorecurrent disease after primary prostate cancer (PCa) treatment. ObjectiveThe aim of this study was to identify the predictors of therapeutic response of MDT for oligorecurrent PCa. Design, setting, and participantsbicentric, retrospective study, including consecutive patients who underwent MDT for oligorecurrent PCa after radical prostatectomy (RP; 2006–2020) was conducted. MDT encompassed stereotactic body radiation therapy (SBRT), salvage lymph node dissection (sLND), whole-pelvis/retroperitoneal radiation therapy (WP[R]RT), or metastasectomy. Outcome measurements and statistical analysisndpoints were 5-yr radiographic progression-free survival (rPFS), metastasis-free survival (MFS), palliative androgen deprivation treatment (pADT)-free survival, and overall survival (OS) together with prognostic factors for MFS following primary MDT. Survival outcomes were studied by Kaplan-Meier survival and univariable Cox regression (UVA). Results and limitationsA total of 211 MDT patients were included; 122 (58%) developed a secondary recurrence. Salvage lymph node dissection was performed in 119 (56%), SBRT in 48 (23%), and WP(R)RT in 31 (15%) of the cases. Two patients received sLND + SBRT and one received sLND + WPRT. Eleven (5%) patients received metastasectomies. The median follow-up since RP was 100 mo, while follow-up after MDT was 42 mo. The 5-yr rPFS, MFS, androgen deprivation treatment(–free survival, castration-resistant prostate cancer–free survival, CSS, and OS after MDT were 23%, 68%, 58%, 82%, 93%, and 87% respectively. There was a statistically significant difference between cN1 (n = 114) and cM+ (n = 97) for 5-yr MFS (83% vs 51%, p < 0.001), pADT-free survival (70% vs 49%, p = 0.014), and CSS (100% vs 86%, p = 0.019). UVA was performed to assess the risk factors (RFs) for MFS in cN1 and cM+. Alpha was set at 10%. RFs for MFS in cN1 were lower initial prostate-specific antigen (PSA) at the time of RP (hazard ratio [95% confidence interval] 0.15 [0.02–1.02], p = 0.053], pN stage at RP (2.91 [0.83–10.24], p = 0.096), nonpersisting PSA after RP (0.47 [0.19–1.12], p = 0.089), higher PSA at primary MDT (2.38 [1.07–5.24], p = 0.032), and number of positive nodes on imaging (1.65 [1.14–2.40], p < 0.01). RFs for MFS in cM+ were higher pathological Gleason score (1.86 [0.93–3.73], p = 0.078), number of lesions on imaging (0.77 [0.57–1.04], p = 0.083), and cM1b/cM1c (non-nodal metastatic recurrence; 2.62 [1.58–4.34], p < 0.001). ConclusionsFollowing MDT, 23% of patients were free of a second recurrence at 5-yr follow-up. Moreover, cM+ patients had significantly worse outcomes in terms of MFS, pADT-free survival, and CSS. The RFs for a metastatic recurrence can be used for counseling patients, to inform prognosis, and potentially select candidates for MDT. Patient summaryIn this paper, we looked at the outcomes of using localized, patient-tailored treatment for imaging-detected recurrent prostate cancer in lymph nodes, bone, or viscera (maximum five recurrences on imaging). Our results showed that targeted treatment of the metastatic lesions could delay the premature use of hormone therapy.

Presence of cells in the polyaneuploid cancer cell (PACC) state predicts the risk of recurrence in prostate cancer.

The nonproliferating polyaneuploid cancer cell (PACC) state is associated with therapeutic resistance in cancer. A subset of cancer cells enters the PACC state by polyploidization and acts as cancer stem cells by undergoing depolyploidization and repopulating the tumor cell population after the therapeutic stress is relieved. Our aim was to systematically assess the presence and importance of this entity in men who underwent radical prostatectomy with curative intent to treat their presumed localized prostate cancer (PCa). Men with National Comprehensive Cancer Network intermediate- or high-risk PCa who underwent radical prostatectomy l from 2007 to 2015 and who did not receive neoadjuvant treatment were included. From the cohort of 2159 patients, the analysis focused on a subcohort of 209 patientsand 38 cases. Prostate tissue microarrays (TMAs) were prepared from formalin-fixed, paraffin-embedded blocks of the radical prostatectomy specimens. A total of 2807 tissue samples of matched normal/benign and cancer were arrayed in nineTMA blocks. The presence of PACCs and the number of PACCs on each core were noted. The total number of cells in the PACC state and the total number of cores with PACCs were significantly correlated with increasing Gleason score (p = 0.0004) and increasing Cancer of the Prostate Risk Assessment Postsurgical(CAPRA-S) (p = 0.004), but no other variables. In univariate proportional hazards models of metastasis-free survival, year of surgery, Gleason score (9-10 vs. 7-8), pathology stage, CAPRA-S, total PACCs, and cores positive for PACCs were all statistically significant. The multivariable models with PACCs that gave the best fit included CAPRA-S. Adding either total PACCs or cores positive for PACCs to CAPRA-S both significantly improved model fit compared to CAPRA-S alone. Our findings show that the number of PACCs and the number of cores positive for PACCs are statistically significant prognostic factors for metastasis-free survival, after adjusting for CAPRA-S, in a case-cohort of intermediate- or high-risk men who underwent radical prostatectomy. In addition, despite the small number of men with complete data to evaluate time to metastatic castration-resistant PCa (mCRPC), the total number of PACCs was a statistically significant predictor of mCRPC in univariate analysisand suggested a prognostic effect even after adjusting for CAPRA-S.

Personalized Prescription of Chemotherapy Based on Assessment of mRNA Expression of BRCA1, RRM1, ERCC1, TOP1, TOP2α, TUBβ3, TYMS, and GSTP1 Genes in Tumors Compared to Standard Chemotherapy in the Treatment of Non-Small-Cell Lung Cancer

Objectives: A growing body of evidence suggests the important role of chemosensitive gene expression in the prognosis of patients with lung cancer. However, studies on combined gene expression assessments for personalized prescriptions of chemotherapy regimens in patients have not yet been conducted. The aim of this work was to conduct a prospective study on the appointment of personalized chemotherapy in patients with non-small-cell lung cancer. Materials and methods: The present study analyzed 85 patients with lung cancer (stage IIB-IIIB). Within this group, 48 patients received individualized chemotherapy, and 37 patients received classical chemotherapy. In the individualized chemotherapy group, the mRNA expression levels of ERCC1, RRM1, TUBB3, TYMS, TOP1, TOP2α, BRCA1, and GSTP1 in lung tissues were measured by quantitative real-time PCR (qPCR), and an individual chemotherapy regimen was developed for each patient according to the results. Patients in the classical chemotherapy group received the vinorelbine/carboplatin regimen. Survival analyses were performed using the Kaplan–Meier method. Prognostic factors of metastasis-free survival (MFS) and overall survival (OS) of patients were identified via Cox’s proportional hazards regression model. Results: MFS and OS were significantly better in the personalized chemotherapy group compared to the classic chemotherapy group (MFS, 46.22 vs. 22.9 months, p = 0.05; OS, 58.6 vs. 26.9 months, p < 0.0001). Importantly, the best metastasis-free survival rates in the group with personalized ACT were achieved in patients treated with the paclitaxel/carboplatin regimen. Based on an assessment of chemosensitivity gene expression in the tumors, the classical chemotherapy strategy also increased the risk of death (HR = 14.82; 95% CI: 3.33–65.86; p < 0.000) but not metastasis (HR = 1.95; 95% CI: 0.96–3.98; p = 0.06) compared to the group of patients with chemotherapy. Conclusions: The use of combined ERCC1, RRM1, TUBB3, TYMS, TOP1, TOP2α, BRCA1, and GSTP1 gene expression results for personalized chemotherapy can improve treatment efficacy and reduce unnecessary toxicity.

Improving locoregional outcome in high-intermediate-risk and high-risk stage I endometrial cancer with surgical staging followed by brachytherapy.

PurposeThis study aims to assess the locoregional efficacy of postoperative vaginal brachytherapy (VBT) alone in patients undergoing surgical staging for early-stage high-intermediate-risk (HIR) and high-risk (HR) endometrial cancer. Materials and MethodsOne hundred and four patients with early-stage HIR and HR endometrial cancer who underwent surgical staging were treated with adjuvant VBT alone. The patients with stage Ib, grade I–III, stage Ia, grade III, lower uterine segment involvement, and lymphovascular invasion (LVI) were included to study. ResultsThe 5- and 10-year overall survival (OS) rates were 87% and 76%, respectively. The 5- and 10-year DFS rates were 86% and 86%, respectively. Among the patients, 92% had endometrioid adenocarcinoma, 2% had undifferentiated carcinoma, 2% had serous papillary carcinoma, and 4% had clear-cell carcinoma. Of the patients, 63% had stage Ib disease, while 37% had stage Ia disease. None of the patients had vaginal or pelvic lymph node recurrence, whereas two had para-aortic lymph node metastasis, one had surgical scar recurrence, one had para-aortic lymph node and brain metastasis, and one had lung metastasis. The presence of lymphatic invasion was found to be a statistically significant prognostic factor for increased distant metastasis rates (p = 0.020). Lymphatic invasion was also regarded as an independent prognostic factor for metastasis-free survival (p = 0.044). ConclusionOur study results suggest that postoperative VBT alone is an effective and safe treatment modality with low complication in patients undergoing surgical staging for HIR and HR endometrial cancer.

Expression of CXCR4 and MMP-2 is associated with poor prognosis in patients with osteosarcoma.

Osteosarcoma is a primary malignant tumor with a high tendency to form metastasis and poor prognosis. Consequently, finding effective early indicators of metastases is crucial for identifying and treating high-risk patients. CXCR4 and MMP-2 have been found to strongly correlate with invasion and metastasis of malignant tumors, including osteosarcoma. Our study evaluated CXCR4 in conjunction with MMP-2 as an important clinicopathological prognostic predictor for metastasis and overall survival of osteosarcoma. 73 patients' clinical data and pathological samples were retrieved for the study. A median time of 36 months follow-up was performed to evaluate for tumor metastasis and patient survival. CXCR4 and MMP-2 proteins in tumor tissues were detected by immunohistochemistry on paraffin-embedded tissue sections. The positive expression rate of CXCR4 and MMP-2 was 68.5% and 54.8% respectively, and of the 45 patients who developed distal metastasis, 33 and 28 patients had positive expression of CXCR4 and MMP-2 respectively. The median metastasis-free survival was 72.00 months in the CXCR4-negative group and 14.00 months in the CXCR4 positive group. Furthermore, median overall survival was 73.77 and 24.00 months in these same two groups. Further, the median metastasis-free survival was 66.51 months in the MMP-2 negative group and 9.00 months in the MMP-2 positive group. The median overall survival was 75.07 and 19.00 months in these same two groups. MMP2 and metastasis remained the significant and independent prognostic factors for metastasis-free survival and overall survival by using the COX regression model adjusted for the multivariate predictors of survival. Our results suggest that metastasis and MMP-2 are both independent prognostic indicators for metastasis-free and overall survival of osteosarcoma patients.

Outcome and Patterns of Relapse in Childhood Parameningeal Rhabdomyosarcoma Treated With Proton Beam Therapy

The standard of care of childhood parameningeal rhabdomyosarcoma (pRMS) is chemotherapy and local radiation therapy. Protons are increasingly being used to decrease late effects. The aim of the present study is to analyze the pattern of relapse and the correlation with dosimetric factors in pRMS treated with proton therapy. This retrospective evaluation includes children treated in our institution for pRMS. Information on demographics, treatment, tumor characteristics, and toxicities and outcome was prospectively collected within the in-house registry. For patients presenting with local relapse, a fusion of the dosimetry with magnetic resonance imaging displaying site and geometry of recurrence was performed. Median follow-up time was 2.9 years (0.5-4.7). Forty-six patients were identified in our institution between July 2013 and November 2017. Main characteristics of patients were as follows: 56.5% male, median age 5.1 years (1.3-17.5), 39.1% alveolar histology, 26.1%, 52.2%, 8.7%, and 13% patients with subgroup risk classification D, E/F/G, H, or metastatic, respectively, median total prescribed dose 55.8 Gy (50.4-56.4). Estimated 2-year local control, metastasis-free survival, event-free survival, and overall survival were 83.8%, 87.8%, 76.9%, and 88.9%, respectively. No acute or late local toxicity exceeding grade 3 was observed. Risk-group was identified as prognostic factor for metastasis-free survival in univariate analysis but not in multivariate analysis (trend: P = .09). In this cohort, dosimetric factors did not correlate with outcome. Isolated local failure happened in 5 of the 11 relapses. Local relapses were matched with dosimetry for 6 patients: 4 of them occurred in the high dose volume and 2 in the intermediate or low dose volume. Proton therapy was effective and well feasible even in a critical cohort. Still, local relapse within the target volume of the radiation therapy remains an important issue in pRMS and new treatment strategies are needed.

Elevated Expression of Transforming Acidic Coiled-Coil Containing Protein 3 (TACC3) Is Associated With a Poor Prognosis in Osteosarcoma.

Transforming acidic coiled-coil containing protein 3 (TACC3) is expressed during the mitotic phase of nuclear division and regulates microtubules. Recently, high TACC3 expression in tumor cells of various cancers including soft tissue sarcoma has been reported. However, its role in osteosarcoma remains unknown. Because we have few prognostic markers for survival in osteosarcoma, we wanted to investigate the potential role of TACC3 in human osteosarcoma and determine if it is associated with survival. (1) Is there a relationship between TACC3 expression and clinicopathologic characteristics such as sex, age (< 20 or ≥ 20 years), histologic type (osteoblastic or others), tumor location (femur or others), American Joint Committee on Cancer staging system (AJCC stage IIA or IIB), tumor necrosis percentage after chemotherapy (< 90% or ≥ 90%), p53 expression (low or high), and Ki-67 expression (low or high)? (2) Is TACC3 expression associated with event-free and overall survival in patients with osteosarcoma? Forty-six conventional patients with osteosarcoma were treated at our institution from 1989 to 2013. Patients were excluded because of unresectable primary site (two patients) and no chemotherapy (two patients). Patients with metastasis at the initial visit (five patients), without pretreatment biopsy samples (two patients), or clinical charts (two patients) were also excluded. The left 33 patients who received neoadjuvant and adjuvant chemotherapy, which consisted of cisplatin/doxorubicin/methotrexate or cisplatin/doxorubicin/methotrexate/ifosfamide, and completed surgical resection with histologic wide tumor margins. Primary tumor samples before chemotherapy were used in this study. We investigated TACC3 expression using immunohistochemical staining and statistically analyzed the TACC3 expression, clinicopathologic characteristics, and event-free and overall survival in patients with osteosarcoma. High TACC3 expression was observed in 19 of 33 osteosarcoma specimens (58%), and this was associated with larger tumor size (ie, AJCC stage IIB in this study; p = 0.002), higher p53 expression (p = 0.007), and higher Ki-67 expression (p = 0.002). The estimated metastasis-free survival at 5 years was 21% (95% confidence interval [CI], 7%-41%) in patients with high TACC3 expression and 79% (95% CI, 47%-93%) in patients with low TACC3 expression (p < 0.001), and the estimated overall survival at 5 years was 34% (95% CI, 13%-56%) in patients with high TACC3 expression and 86% (95% CI, 54%-96%) in patients with low TACC3 expression (p < 0.001). Furthermore, high TACC3 expression was an independent poor prognostic factor for metastasis-free survival with a hazard ratio of 3.89 (95% CI, 1.07-19.78; p = 0.039) as well as overall survival with 4.41 (95% CI, 1.01-32.97; p = 0.049). High TACC3 expression was associated with aggressive clinicopathologic features and unfavorable prognosis in these patients with osteosarcoma. Our preliminary results suggest that further analysis about mutation or an inactive form of TACC3 would be useful to understand the mechanism of abnormal TACC3 expression in patients with osteosarcoma. If these findings are substantiated in larger studies, TACC3 might be useful for predicting survival and a potential therapeutic target for osteosarcoma. Level III, therapeutic study.

Abstract 2530: Validation and pathway analysis of a metastasis-specific microRNA signature in primary colon cancer

Abstract Introduction: In lymph node negative (LNN) colon cancer 20% of the patients develops disease recurrence. Identification of these patients is needed. MicroRNAs (MiRNAs), a group of short non-coding RNAs, can function as tumor suppressors or oncogenes. Hur et al recently found 6 differentially expressed MiRNAs when comparing primary colorectal cancer and matched liver metastasis (MiR-320, MiR-221, MiR-30b, MiR-10b, MiR-885-5p, Let-7i). The expression of 2 MiRNAs was significantly correlated with distant metastasis (low Let-7i expression and high MiR-10b) in primary colorectal cancers. When split at the median and combined into a signature (Let-7i high and MiR-10b low [n=22] vs. Let-7i low and/or MiR-10b high[n=122]), the first group showed 100% metastasis free survival (MFS). We assessed the prognostic value of these MiRNAs and the signature in a clinically well-defined cohort of primary colon cancers. Methods: Expression of the 6 MiRNAs were measured using RT-qPCR in a cohort of 232 colon cancer patients (n=155 untreated LNN and n=77 adjuvant treated lymph node positive [LNP] patients) selected from the MATCH-cohort. Expression levels were related to microsatellite instability (MSI), and MFS, hepatic metastasis free (HFS) and overall survival (OS). Results: MiR-221, Mir-30b, Mir-10b and MiR-885-5p levels were significantly associated with MSI. In univariate Cox regression analysis, MiR-30b was significantly associated with MFS (HR=1.23 p=0.004), and MiR-30b and Let-7i were associated with HFS (HR=2.52 p=0.005 and HR=0.40 p&amp;lt;0.001, respectively) in the total group. In the LNN group results were similar for MiR-30b (MFS HR=2.07 p=0.008 and HFS HR=2.94 p=0.005) and Let-7i (HFS HR=0.28 p&amp;lt;0.001), but did not reach significance in the LNP group. These associations remained significant when correcting for MSI. Pathway analysis for MiR-30b and Let-7i with the GSEA hallmark gene sets between the 50 samples with the highest and 50 samples with lowest expression revealed a significantly higher expression of the TGF-beta pathway in the MiR-30b high group, and a significantly higher expression of the EMT pathway in the Let-7i high group. MiR-30b and Let-7i expression was split at the median level and combined into two groups (‘Let-7i high and MiR-30b low’ vs. ‘Let-7i low and/or MiR-30b high’). The ‘Let-7i high and MiR-30b low’ group in the total group (n=74) had a significantly better 5-yr HFS (100% vs 87.4% p=0.002), and had a significantly better 5-yr MFS (91.7% vs 78% p=0.036) and HFS (100% vs 87.8% p=0.01) in the LNN group (n=52). Conclusion: in our cohort and more specifically the LNN group, we confirmed Let-7i and identified MiR-30b as a prognostic factor for MFS and HFS. We did not confirm the prognostic value of Mir-10b. The combination of Let-7i and MiR-30b identified a group with a 100% HFS. Pathway analysis showed higher expression of the TGF-beta pathway in the MiR-30b high group, and higher expression of the EMT pathway in the Let-7i high group. Citation Format: Robert R. Coebergh van den Braak, Anieta M. Sieuwerts, Zarina S. Lalmahomed, Marcel Smid, Vanja de Weerd, Michelle van der Vlugt - Daane, Anne van Galen, Shanshan Xiang, Katharina Biermann, John A. Foekens, John W. Martens, Jan N. IJzermans. Validation and pathway analysis of a metastasis-specific microRNA signature in primary colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2530. doi:10.1158/1538-7445.AM2017-2530

Expression and prognostic significance of PDGF ligands (A, B, C, and D) and PDGFR (A, B, and L) in soft-tissue sarcomas and GIST.

11067 Background: Sarcomas are a variety of rare connective tissue cancers. Doxorubicin and olaratumab (Ab against PDGFRA) improved survival in a recent Phase 1/2 study. Besides PDGFRA mutated gastrointestinal stroma tumor (GIST) and dermatofibrosarcoma protuberans, the role of PDGFR and the according ligands in the biology of sarcoma remain unclear. Methods: The expression levels of PDGF (A,B,C,D) and PDGFRs (A,B,L) were studied in a series of 255 sarcoma pts in localized phase using the Agilent 014850 platform. Data are available online (http://atg-sarc.sarcomabcb.org). Histologies were GIST (n = 60), myxoid liposarcoma (MLPS, n = 50), synovial sarcoma (SyS, n = 58), and sarcoma with complex genomics (SCG, n = 87). Expression levels were analyzed and tested for prognostic values for metastasis free survival (MFS) in uni- and multivariate analysis using SPSS 19.0. Results: Expression levels (ELs) of PDGFs and PDGFRs varied across histotypes: PDGFA levels were highest in SyS and lowest in MLPS (p &lt; 0.0001). PDGFB and C levels were lower in GIST (p &lt; 0.0001), while PDGFD ELs were similar across histological subtypes. PDGFRA ELs were highest in MLPS, while PDGFRB &amp; L ELs were lowest in GIST and SyS (p &lt; 0.0001 all). Complex patterns of correlation of expression between ligands and receptors were observed in each individual subtypes. PDGFA ELs above median were associated with a marginally higher risk of metastasis . Conversely, PDGFD ELs above median was associated with a reduced risk of metastasis in the whole cohort (p = 0.02). The ELs of the 3 receptors were not correlated to MFS. In multivariate analysis using Cox model on the non-GIST sarcoma cohort (histology, grade, depth, with size, PDGFA, PDGFD as continuous variables): histology, size, grade and PDGFA ELs were independent adverse prognostic factors (PF), while PDGFD ELs was a favorable PF for MFS. In the GIST cohort, testing AFIP score, PDGFA &amp; D ELs as continuous variable, PDGFD ELs was also an independent favorable PF for MFS, in addition to AFIP score. Conclusions: The expression of PDGFs and the according receptors varies across sarcoma histological subtypes. PDGFA and D expression levels correlate independently to the risk of metastatic relapse.

Prognostic significance of chemotherapy-induced necrosis in osteosarcoma patients receiving pasteurized autografts.

BackgroundAmong various reconstruction methods after wide excision for osteosarcoma, pasteurized autograft is often preferred. While the whole area of the tumor can be assessed for chemotherapy-induced necrosis, one of the important prognostic factors, in other reconstructive techniques, only a portion removed from a wide-resection specimen is available when using pasteurized autograft method. The assessment, therefore, may be unreliable. We analyzed the prognostic significance of the chemotherapy-induced necrosis in osteosarcoma patients who underwent reconstruction with pasteurized autografts.Patients and methodsWe reviewed the records of osteosarcoma patients who underwent treatment in our institution from 1998 to 2013. Cases of reconstruction with pasteurized autografts were defined as the patient group, and the same number of patients who underwent other reconstruction methods served as controls. Chemotherapy-induced necrosis was evaluated for removed extra-osseous and curetted intramedullary tumor tissues.ResultsA total of 22 patients were identified; the median age was 15.5 years, and there were 12 males. The most common tumor location was the distal femur. The most common histological subtype was osteoblastic. Median size was 8.1 cm. Disease status was stage IIB in 13 patients and IIA in 9. Median follow-up was 76 months. No differences between the patient and control groups were observed in potential prognostic factors, overall survival, metastasis-free survival, or recurrence-free survival. Univariate analyses demonstrated that histological response was a significant prognostic factor for metastasis-free survival and also significant for recurrence-free survival.ConclusionChemotherapy-induced necrosis grading, using only available tumor tissues, could be a prognostic factor for osteosarcoma patients receiving pasteurized autografts for reconstructive surgery.

Combined evaluation of LC3B puncta and HMGB1 expression predicts residual risk of relapse after adjuvant chemotherapy in breast cancer

In spite of adjuvant chemotherapy, a significant fraction of patients with localized breast cancer (BC) relapse after optimal treatment. We determined the occurrence of cytoplasmic MAP1LC3B/LC3B (microtubule-associated protein 1 light chain 3B)-positive puncta, as well as the presence of nuclear HMGB1 (high mobility group box 1) in cancer cells within surgical BC specimens by immunohistochemistry, first in a test cohort (152 patients) and then in a validation cohort of localized BC patients who all received adjuvant anthracycline-based chemotherapy (1646 patients). Cytoplasmic LC3B+ puncta inversely correlated with the intensity of SQSTM1 staining, suggesting that a high percentage cells of LC3B+ puncta reflects increased autophagic flux. After setting optimal thresholds in the test cohort, cytoplasmic LC3B+ puncta and nuclear HMGB1 were scored as positive in 27.2% and 28.6% of the tumors, respectively, in the validation cohort, while 8.7% were considered as double positive. LC3B+ puncta or HMGB1 expression alone did not constitute independent prognostic factors for metastasis-free survival (MFS) in multivariate analyses. However, the combined positivity for LC3B+ puncta and nuclear HMGB1 constituted an independent prognostic factor significantly associated with prolonged MFS (hazard ratio: 0.49 95% confidence interval [0.26–0.89]; P = 0.02), and improved breast cancer specific survival (hazard ratio: 0.21 95% confidence interval [0.05–0.85]; P = 0.029). Subgroup analyses revealed that within patients with poor-prognosis BC, HMGB1+ LC3B+ double-positive tumors had a better prognosis than BC that lacked one or both of these markers. Altogether, these results suggest that the combined positivity for LC3B+ puncta and nuclear HMGB1 is a positive predictor for longer BC survival.

The Preoperative AST/ALT (De Ritis) Ratio Represents a Poor Prognostic Factor in a Cohort of Patients with Nonmetastatic Renal Cell Carcinoma

Aminotransaminases, which are strongly involved in cellular metabolism and cancer cell turnover, represent easily measureable, potential blood based biomarkers. We evaluated the prognostic value of the preoperatively assessed AST/ALT (De Ritis) ratio on clinically meaningful end points in a large European cohort of patients with nonmetastatic renal cell carcinoma. We retrospectively evaluated clinicopathological data on 698 patients with nonmetastatic renal cell carcinoma operated on between 2005 and 2013 at a single tertiary academic center. The potential prognostic value of the AST/ALT ratio was analyzed using the Kaplan-Meier method, and univariate and multivariate Cox proportional regression models. The impact of the ratio on the predictive accuracy of the Leibovich prognosis score was determined by the Harrell c-index. An increased (1.26 or greater) preoperative AST/ALT ratio was statistically significantly associated with several well established prognostic factors, including pathological T stage, as well as with histological tumor necrosis (p <0.05). On multivariate analysis an increased preoperative AST/ALT ratio was an independent prognostic factor for metastasis-free survival (HR 1.61, 95% CI 1.25-2.07, p <0.001) and overall survival (HR 1.76, 95% CI 1.34-2.32, p <0.001). The Harrell c-index was 0.77 using the Leibovich prognosis score and 0.81 when AST/ALT was added. In our study cohort with nonmetastatic renal cell carcinoma the preoperatively assessed AST/ALT ratio represented an independent prognostic factor. This ratio might further improve the predictive accuracy of well established prognosis scores.

Expression of the hypoxia-inducible monocarboxylate transporter MCT4 is increased in triple negative breast cancer and correlates independently with clinical outcome

Background18Fluor-deoxy-glucose PET-scanning of glycolytic metabolism is being used for staging in many tumors however its impact on prognosis has never been studied in breast cancer. MethodsGlycolytic and hypoxic markers: glucose transporter (GLUT1), carbonic anhydrase IX (CAIX), monocarboxylate transporter 1 and 4 (MCT1, 4), MCT accessory protein basigin and lactate-dehydrogenase A (LDH-A) were assessed by immunohistochemistry in two cohorts of breast cancer comprising 643 node-negative and 127 triple negative breast cancers (TNBC) respectively. ResultsIn the 643 node-negative breast tumor cohort with a median follow-up of 124months, TNBC were the most glycolytic (≈70%), followed by Her-2 (≈50%) and RH-positive cancers (≈30%). Tumoral MCT4 staining (without stromal staining) was a strong independent prognostic factor for metastasis-free survival (HR=0.47, P=0.02) and overall-survival (HR=0.38, P=0.002). These results were confirmed in the independent cohort of 127 cancer patients. ConclusionGlycolytic markers are expressed in all breast tumors with highest expression occurring in TNBC. MCT4, the hypoxia-inducible lactate/H+ symporter demonstrated the strongest deleterious impact on survival. We propose that MCT4 serves as a new prognostic factor in node-negative breast cancer and can perhaps act soon as a theranostic factor considering the current pharmacological development of MCT4 inhibitors.

Extracellular proteases in renal cell carcinoma.

363 Background: Tumor growth and metastasis relies on extracellular matrix remodeling, which is primarily mediated by the matrix metalloproteinase (MMP) family of enzymes. Extracellular matrix metalloprotease inducer (EMMPRIN, E) is a glycoprotein also implicated in tumor progression and has been shown to function in concert with MMPs. MMPs and E are highly expressed on the surface of many malignant tumors, including renal cell carcinoma (RCC). We investigated whether MMPs and E represent molecular prognostic factors in RCC. Methods: Protein expression was evaluated in 77 patients with immunohistochemical analysis of a tissue microarray (TMA) consisting of two tumor samples and an adjacent normal renal sample from each patient. Tumors represented on the TMA were clear cell (57), chromophobe (9), papillary (4), oncocytomas (3), clear cell with sarcomatoid differentiation (2), and metastatic (2). Protein expression scores were correlated with clinical outcomes to determine prognostic significance by univariate and multivariate analyses. Results: Using univariate analysis of all RCC tumors, membranous E correlated with overall survival (OS), metastasis-free survival (MFS), size, stage and overall 5-yr survival. Cytoplasmic E correlated with OS, MFS, size, grade, stage and 5-yr survival. Membranous MT1-MMP correlated with MFS, and endothelial MMP-2 correlated with grade. In multivariate analysis, membranous E was an independent indicator of stage, whereas cytoplasmic E was an independent indicator for size, stage, grade and MFS. In the clear cell population, membranous MT1-MMP correlated with OS and MFS, and endothelial MMP-2 correlated with grade in univariate analysis. In multivariate analysis, membranous MT1-MMP was an independent indicator for MFS. Conclusions: We identified EMMPRIN as an independent prognostic indicator for size, grade, stage, and MFS in RCC. While MT1-MMP and MMP-2 both significantly correlated with disease outcome in the clear cell RCC population, only MT1-MMP was an independent prognostic factor for MFS. We conclude that MMPs and E mediate important steps in RCC progression. Further investigation is warranted using larger datasets and a prospective study to determine the strength of the prognostic significance of these findings. No significant financial relationships to disclose.

Impact of reducing clinical target volume on efficacy of intensity modulated radiation therapy for nasopharyngeal carcinoma

Objective To evaluate the impact of reducing clinical target volume (CTV) on the efficacy of intensity modulated radiation therapy (IMRT) for nasopharyngeal carcinoma (NPC) . Methods Between August 2003 and March 2007, 380 NPC patients were treated with IMRT with reduced CTV.CTV1, defined as high risk region, included GTV +5 - 10 mm margin and the entire nasopharyngeal mucosa +5 mm submucosal volume; CTV2, designed for potentially involved regions, included the nasopharyngeal cavity (limited to the posterior part of nasal cavity only), maxillary sinus (limited to 5 mm anterior to the posterior nasal aperture and maxillary mucosa), pterygopalatine fossa, posterior ethmoid sinus,parapharyngeal space, skull base, anterior third of clivus and cervical vertebra, inferior spheniod sinus and cavernous sinus and internal group of retropharyngeal lymph nodal regions from the base of skull to cranial edge of the second cervical vertebra. The prescription dose was: GTV 66. 00 -69. 75 Gy/30 - 33 f, CTV1 60. 00 -66. 65 Gy,CTV2/CTVN 54. 0 -55.8 Gy. 308 patients with stage Ⅲ or Ⅳ diseases also received cisplatin-based neoadjuvant chemotherapy. Results The follow-up rate was 100%. 145 patients were followed-up to 3 years. The 3-year estimated local control, regional control, metastasis-free survival,disease-free survival and overall survival rates were 94. 9%, 97.4%, 86. 2%, 80. 9% and 89. 0%,respectively. Multivariate analysis revealed that N-classification was a significant prognostic factor for metastasis-free survival (x2 = 20. 80, P = 0. 001), N-classification (x2 = 18. 30, P = 0. 003) and age (x2 =7. 31, P =0. 004) were independent prognostic factors for overall survival. Grade 2 xerostomia was observed in 5.6% of the patients after two years of IMRT, no Grade 3 or 4 xerostomia was observed. Local, regional,and distant failures were developed in 4. 2%, 2. 6% and 12. 1% of the patients, respectively. Conclusions The IMRT approach with reduced CTV2 provids a favorable outcome for NPC with acceptable toxicities. Key words: Nasopharyngeal neoplasms/radiotherapy; Radiotherapy, intensity-modulated; Clinical target volume; Prognosis

Long-term outcomes of eye-conserving treatment with Ruthenium 106 brachytherapy for choroidal melanoma

Purpose To evaluate long-term outcomes of eye-conserving treatment using Ruthenium-106 plaque brachytherapy with or without transpupillary thermotherapy (TTT) for small to intermediate size choroidal melanomas. Methods Outcomes of 425 consecutive patients were analysed. The median basal tumour diameter was 10.9 mm (range 4.8–15.9 mm), and the median apical height 4.2 mm (range 1.2–9.3 mm). Brachytherapy doses ranged from 400 to 600 Gy with TTT (86%), or from 600 to 800 Gy without TTT (14%), specified at the scleral surface. Kaplan–Meier survival curves, log-rank tests and Cox regression analysis were used for analysis. Results Median follow-up was 50 months. Five-year actuarial local control was 96%. Five-year overall and metastases-free survival rates were 79.6% and 76.5%. Prognostic factors for metastasis-free survival were peripheral location ( p = 0.02) and smaller basal diameter ( p < 0.001). No dose effect relationships were found. Radiation side effects were frequent, with 2- and 5-year rates free of radiation complications of 60% and 35%. Five-year enucleation rate was 4.4% (10 for local recurrence, 7 for complications). Cosmetic and functional (visual acuity >0.10) eye preservation rates were 96% and 52% at 5 years. Conclusions Ruthenium-106 brachytherapy for choroidal melanoma provides excellent rates of local control and eye preservation.

Abstract 3668: Relevance of BCAR4 in tamoxifen resistance and tumor aggressiveness of breast cancer

Abstract Background: Tamoxifen is effective in the treatment of estrogen receptor alpha (ER)-positive breast cancer. However, almost all responding patients develop resistance. Understanding the mechanisms leading to resistance is crucial for the development of more efficient therapies. In a functional screen for genes causing tamoxifen resistance, we have identified the breast cancer antiestrogen resistance 4 (BCAR4) gene. Forced expression of BCAR4 in estrogen-dependent breast cancer cell lines induced antiestrogen resistance. Objective: We explored whether BCAR4 mRNA levels in primary tumors predict tamoxifen resistance and/or tumor aggressiveness, and searched for signaling pathways involved in BCAR4-induced resistance to tamoxifen. Experimental design: BCAR4 mRNA levels were measured by quantitative RT-PCR. In 280 ER-positive primary tumors from patients receiving tamoxifen as first-line monotherapy for advanced disease, the associations between BCAR4 mRNA levels and progression-free survival (PFS) and clinical benefit were evaluated. BCAR4 mRNA levels in primary tumors from 506 patients with lymph node-negative, ER-positive cancer were evaluated for their association with distant metastasis-free survival (MFS). None of them received systemic adjuvant therapy, thereby preventing confounding effects of systemic treatment. In order to study BCAR4 function, we forced the expression of BCAR4 in estrogen-dependent ZR-75-1 breast cancer cells, and analyzed changes in phospho-protein expression. Results: In the study for the association of BCAR4 with tamoxifen resistance, high BCAR4 mRNA levels were significantly associated with a shorter PFS and poor clinical benefit (measurable tumor response or no change &amp;gt; 6 months). Multivariate analysis including the traditional predictive factors indicated that a high BCAR4 mRNA level is an independent predictive factor for PFS. In the assessment of tumor aggressiveness, high BCAR4 mRNA levels were associated with a shorter MFS and overall survival (OS). Multivariate analyses showed that a high BCAR4 mRNA level was an independent prognostic factor for MFS and OS. Forced expression of BCAR4 induced phosphorylation of ERBB2, ERBB3, and their down-stream mediators ERK1/2 and AKT in ZR-75-1 cells. Proliferation of these cells was inhibited by the knockdown of ERBB2 or ERBB3 with specific siRNAs, confirming the role of these receptors in BCAR4-induced tamoxifen resistance. The knockdown of ER had no effect on the proliferation of BCAR4-expressing cells, indicating that this mechanism of resistance is independent of ER signaling. Conclusions: Our data indicate that high BCAR4 mRNA levels predict resistance to tamoxifen treatment and poor outcome. The results of our experimental cell model also suggest that tumors expressing BCAR4 may rely on ERBB2/ERBB3 signaling. Therefore, patients with tumors expressing BCAR4 may be eligible to receive ERBB-targeted therapy. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3668.