Total Tumor Area Research Articles

PD71-11 P53 FOCAL PROTEIN EXPRESSION IN PRIMARY PROSTATE TUMORS AND LYMPHATIC VESSEL INVASION PREDICT BIOCHEMICAL RECURRENCE AND METASTATIC PROGRESSION

You have accessJournal of UrologyProstate Cancer: Markers III1 Apr 2017PD71-11 P53 FOCAL PROTEIN EXPRESSION IN PRIMARY PROSTATE TUMORS AND LYMPHATIC VESSEL INVASION PREDICT BIOCHEMICAL RECURRENCE AND METASTATIC PROGRESSION William Gesztes, Jennifer Cullen, Denise Young, Yongmei Chen, Allen Burke, Albert Dobi, Gyorgy Petrovics, Inger Rosner, Shiv Srivastava, and Isabell Sesterhenn William GesztesWilliam Gesztes More articles by this author , Jennifer CullenJennifer Cullen More articles by this author , Denise YoungDenise Young More articles by this author , Yongmei ChenYongmei Chen More articles by this author , Allen BurkeAllen Burke More articles by this author , Albert DobiAlbert Dobi More articles by this author , Gyorgy PetrovicsGyorgy Petrovics More articles by this author , Inger RosnerInger Rosner More articles by this author , Shiv SrivastavaShiv Srivastava More articles by this author , and Isabell SesterhennIsabell Sesterhenn More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.3178AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES p53 has been widely studied in prostate cancer (CaP). Next-Gen Sequencing (NGS) studies of prostate tumors reveal that p53 mutations are frequently observed in castration resistant CaP genomes. Recently, Haffner et al. (JCI, 2013) reported data on a single patient with a p53 mutation in a primary, well differentiated tumor at time of radical prostatectomy (RP) that was linked to future metastatic lesions. Lymphatic vessel invasion (LI) has also been demonstrated to predict poor CaP outcomes. The study purpose was to evaluate independent and combined roles of p53 and LI status in predicting CaP progression in a RP patient cohort with long-term follow-up. METHODS This retrospective RP cohort study was comprised of CaP patients (50 metastatic; 138 randomly sampled, non-metastatic) enrolled at the Walter Reed National Military Medical Center between 1993 and 2013. Representative sections of whole mounted RP specimens were examined. LI and p53 status were assessed using immunohistochemistry staining with Biocare Medical D2-40 & p53 Tumor Suppressor Protein monoclonal antibodies. In CaP index tumors, p53 status was evaluated as percent p53 positive (+) tumor cells divided by total index tumor area (0%, 1-5%, >5%). Kaplan Meier (KM) estimation curves were used to examine time to biochemical recurrence (BCR) and distant metastasis as a function of p53 and LI status. RESULTS Of the 188 eligible patients, median age at RP and follow-up time were 60.4 and 12.7 years, respectively. Nearly half (49.2%) of tumors stained focally as p53+ while 26.6% of patients had evidence of LI. p53+ patients had higher pathologic T stage (61.6% p53+ vs. 38.4% p53-, p = 0.0068). In KM models, p53+ >5% alone predicted BCR (p = 0.0075) and distant metastasis (p = 0.0053). When p53 status was examined in combination with LI, the poorest CaP outcomes were observed for those jointly LI+/p53 >5% (Figure 1a-b, p < 0.0001). CONCLUSIONS p53 protein expression, alone and in combination with LI status, predicts CaP progression after RP. These data suggest that determination of p53 alterations is warranted for improved prognostication of CaP progression. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e1359-e1360 Advertisement Copyright & Permissions© 2017MetricsAuthor Information William Gesztes More articles by this author Jennifer Cullen More articles by this author Denise Young More articles by this author Yongmei Chen More articles by this author Allen Burke More articles by this author Albert Dobi More articles by this author Gyorgy Petrovics More articles by this author Inger Rosner More articles by this author Shiv Srivastava More articles by this author Isabell Sesterhenn More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

T cell infiltration in matched renal biopsy (bx) and nephrectomy (nx) samples in renal cell carcinoma (RCC).

472 Background: T cell infiltration in tumors has been investigated as a biomarker of response to checkpoint inhibitors. A neo-adjuvant trial of checkpoint inhibition in locally-advanced RCC is ongoing at Cleveland Clinic, where T cell infiltration in pre-treatment renal mass bx will be compared to post-treatment nx specimens. However, there are no data regarding the association of T cell infiltration in matched bx and nx samples without intervening treatment. Understanding this association will enable further study of this potential biomarker in future neo-adjuvant studies. Methods: Matched bx and nx samples (without intervening systemic therapy) were identified from patients with non-metastatic RCC. Demographic and pathological data were collected from chart review. Selected tissue sections from bx and nx samples of each patient were reviewed, and marked for tumor and intra-tumoral lymphocytes by the pathologist. Immunohistochemistry (IHC) was utilized to stain these selected tissue sections for T cell markers (CD3, CD4 and CD8). Intra-tumoral T cells were then quantified in the pre-marked tissue sections as counts per total tumor area surveyed, using Image-Pro Plus (Media Cybernetics, Inc.). Spearman correlation (ρ) was used to measure the strength of association of T cell infiltration between matched samples. Results: 30 matched pairs were investigated. The median interval between bx and nx was 2.8 (0.2-87.7) months. Clear cell was the most common histology (29/30; 97%). 15/30 (50%) had grade 3-4 tumors, 2/19 (11%) patients had sarcomatoid features, 7/25 (29%) had necrosis, and 8/28 (29%) had lymphovascular invasion. We found a positive correlation between the frequencies of CD8+ T cells between matched bx and nx samples (ρ= 0.39; p=0.03). CD3+ and CD4+T cells did not show significant correlation. (Table) Conclusions: Bx material can be used to accurately assess the degree of CD8+T cell infiltration in RCC. [Table: see text]

P3.03-037 Impact of Sarcomatoid Component in Patients with Biphasic Mesothelioma: Review of 118 Patients: Topic: Mesothelioma Clinical

Biphasic mesothelioma has a poor prognosis. There is no clear evidence on the role of multimodality treatment in patients with biphasic mesothelioma. The aim of this study was to analyze the impact of pathological features on survival, to determine which patients may benefit from multimodality treatment. Between January 2005 and December 2015, 214 patients with biopsy-proven biphasic mesothelioma were retrospectively identified to fulfil our inclusion criteria. The primary outcome was survival measured from time of diagnosis. Two slides were reviewed for each patient by a specialist thoracic pathologist. Slides were stained with Hematoxylin and Eosin (H+E) and the immunohistochemically-stained slides were digitally scanned and analyzed using a Hamamatsu Nanozoomer scanner (Hamamatsu ‘NDP.View2’). The proportion of epithelioid and sarcomatoid components on each slide was mapped and its area in mm2 recorded as a percentage of the total tumor area studied. Necrosis and lymphovascular invasion were analyzed. Patients with no slides available were excluded from the analysis (n=96). All eligible patients (n=118) were followed up until May 2016. One hundred and eighteen patients were included in the analysis, 106 (89.9%) were male with a median age of 73 (range 53 - 91). Twenty-eight patients (23.7%) underwent Pleurectomy Decortication (PD) and 90 patients received medical treatment alone, either with chemotherapy or best supportive care. The median overall survival (OS) was 11.2 months (range 0.3 – 36.2). At 1 year and 2 years, 49.1% and 6.4% of patients were alive respectively. Uni-variable analysis revealed both age and PD to be associated with improved survival (p=0.004 and p=0.004). Patients treated with PD had OS of 12.8 months (range 5.6 - 36), compared to 9.2 months (range 0.3 – 31.8) in patients receiving medical treatment alone. No lymphovascular invasion was identified in any specimen. Necrosis was not correlated with survival (p=0.76). The proportion of epithelioid (p=0.45) or sarcomatoid (p=0.60) component within the specimen did not correlate significantly with overall survival. This remained true when patients undergoing surgical PD (epitheloid p=0.42 and sarcomatoid p=0.60) and medical treatment (epitheloid p=0.43 and sarcomatoid p=0.11) were analyzed as separate subgroups. The prognosis for patients with biphasic mesothelioma remains poor, even after multimodality treatment including pleurectomy decortication (PD). However, necrosis and the proportion of sarcomatoid histology is not helpful in selecting patients with more favorable prognosis, who may benefit from a multimodality approach.

Abstract B80: Stroma in pancreatic cancer: Automated quantification and its prognostic significance

Abstract Tumor stroma (TS) is a cardinal component of pancreatic ductal adenocarcinoma (PDAC); it comprises 60 to 90% of tumor volume and is an emerging treatment target. There are conflicting reports of the role that TS plays in disease progression and its prognostic significance. The objective of this study was to develop a high-throughput, operator independent method for objectively quantifying TS density (TSD) and to evaluate the prognostic significance of TSD in resected tumors. Tissue sections of primary tumor from 126 patients undergoing pancreaticoduodenectomy for PDAC from 1979 to 2008 and followed by observation only were stained with hematoxylin and eosin (H&amp;E). Sections were digitally uploaded and location of tumor was annotated using ImageScope. Annotated images were imported to Definiens Architect. By training on representative areas, an algorithm was created to segment the tissue into different regions-of-interest (ROI): tumor epithelium, TS, and inflammatory infiltrate. Next, the algorithm was used to quantify the above described ROIs in each tissue section. TSD was calculated as tumor stroma area over total tumor area. A cox proportional hazards model was used to compare overall survival (OS) and recurrence free survival (RFS) vs TSD controlling for age, sex, tumor grade, T, N, and overall stage, smoking status, and American Society of Anesthesia (ASA) physical classification score. Median TSD was 0.637. High TSD was significantly associated with RFS (p=0.043) and OS (p=0.037). In the low TSD group, median RFS and OS was 8.9 months and 11.5 months, respectively. In the high TSD group, median RFS and OS was 13.9 and 18.3 months, respectively. In this cohort, 18 patients were identified with an OS greater than 5 years. This cohort had a higher median TSD of 0.71. After excluding those with a survival greater than 5 years, there was still a trend toward high TSD and worse OS (p=0.052). Our novel automated algorithm for quantification of TSD allows for high throughput, reproducible, and objective analysis of H&amp;E stained PDAC slides. The median TSD in our study obtained using our novel algorithm was 0.637, in agreement with prior manual descriptions of PDAC tumors having a TSD of greater than 60%. We also found high TSD to be a positive independent prognostic variable in resected tumors in patients not receiving adjuvant therapy. Further understanding of the balance of TS in restraining tumor progression vs impeding chemotherapeutic delivery is needed to successfully manipulate stroma in the treatment of PDAC. Citation Format: Robert J. Torphy, Rashid Naim, Bentley Midkiff, Keith Volmar, Eric Collisson, Jen Jen Yeh.{Authors}. Stroma in pancreatic cancer: Automated quantification and its prognostic significance. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B80.

Abstract A109: Radium-223 dichloride inhibits tumor growth and tumor-induced bone growth in osteoblastic prostate cancer models

Abstract Radium-223 dichloride (radium-223), an alpha particle-emitting calcium-mimetic, improves overall survival in prostate cancer patients with symptomatic bone metastases. We have defined radium-223 mode-of-action and efficacy in two clinically relevant prostate cancer xenograft models demonstrating PSA expression and osteoblastic growth upon intratibial inoculation of cancer cells. Immunocompromised male mice were inoculated with human LNCaP or patient-derived LuCaP 58 prostate cancer cells in the intratibial compartment and subsequently stratified into treatment groups based on lesion grade and/or serum PSA levels. Radium-223 (300 kBq/kg) or vehicle was administered intravenously, two times at 4-week intervals during the experiment. X-rays and serum samples were obtained biweekly and at sacrifice. Soft tissue tumors were examined macroscopically at sacrifice and tissue samples were collected and processed for γ-counter measurements, micro-CT, autoradiography and histology. Radium-223 treatment inhibited tumor-induced osteoblastic bone growth as indicated by reduced bone volume and surface in LNCaP and LuCaP 58 prostate cancer mouse models. In addition, radium-223 treatment suppressed metabolic activity in bone as evidenced by decreased number of osteoblasts and osteoclasts relative to bone surface and reduced levels of the bone formation marker PINP. Radium-223 resulted in lower PSA values as early as two weeks after the first dose, indicating constrained tumor growth following treatment. This phenomenon was further supported by reduced total bone lesion tissue and tumor area in LNCaP and LuCaP 58 models and increased percentage of necrotic tumor area in the LuCaP 58 model in radium-223-treated mice as compared to vehicle-treated mice. Moreover, DNA double-strand breaks were increased in cancer cells 24 hours post radium-223 treatment in the LuCaP 58 model providing further evidence of anti-tumor effects. Radium-223-treated mice exhibited less visceral metastases in the LuCaP 58 model (not significant). Based on autoradiography, radium-223 was deposited in the intratumoral bone matrix and in conjunction with osteoblasts in osteoblastic metastases. Our results demonstrate that radium-223 dichloride is successfully incorporated into the intratumoral bone matrix and inhibits tumor growth in both cell line- and patient-derived osteoblastic prostate cancer metastasis models. Given the α-particle range of 50-80 μm, potent radiation effects on the tumor microenvironment are evident whereas relevant effects on the more distant bone marrow are not expected. Taken together, radium-223 therapy exhibits a dual mode-of-action that impacts tumor growth and tumor-induced bone reaction, both important players in the destructive vicious cycle of osteoblastic bone metastasis in prostate cancer. Citation Format: Mari I. Suominen, Katja M. Fagerlund, Dominik Mumberg, Karl Ziegelbauer, Sanna-Maria Käkönen, Jussi M. Halleen, Robert L. Vessella, Arne Scholz. Radium-223 dichloride inhibits tumor growth and tumor-induced bone growth in osteoblastic prostate cancer models. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr A109.

Abstract 3447: Radium-223 dichloride exhibits dual mode-of-action inhibiting both tumor and tumor-induced bone growth in two osteoblastic prostate cancer models

Abstract Radium-223 dichloride (radium-223), an alpha particle-emitting calcium-mimetic, improves overall survival in prostate cancer patients with symptomatic bone metastases. Here, we define radium-223 mode-of-action and efficacy in two clinically relevant prostate cancer xenograft models demonstrating PSA expression and osteoblastic growth upon intratibial inoculation of cancer cells. Immunocompromized male mice were inoculated with human LNCaP or patient-derived LuCaP 58 prostate cancer cells in the intratibial compartment and subsequently stratified into treatment groups based on lesion grade and/or serum PSA levels. Radium-223 (300 kBq/kg) or vehicle was administered intravenously, two times at 4-week intervals during the experiment. X-rays and serum samples were obtained biweekly and at sacrifice. Soft tissue tumors were examined macroscopically at sacrifice and tissue samples were collected and processed for γ-counter measurements, micro-CT, autoradiography and histology. Radium-223 treatment inhibited tumor-induced osteoblastic bone growth as indicated by reduced bone volume and surface in LNCaP and LuCaP 58 prostate cancer mouse models. In addition, radium-223 treatment suppressed metabolic activity in bone as evidenced by decreased number of osteoblasts and osteoclasts relative to bone surface and reduced levels of the bone formation marker PINP. Radium-223 resulted in lower PSA values as early as two weeks after the first dose, indicating constrained tumor growth following treatment. This phenomenon was further supported by reduced total tissue and tumor area in tibia in LNCaP and LuCaP 58 models and increased percentage of necrotic tumor area in the LuCaP 58 model in radium-223-treated mice as compared to vehicle-treated mice. Moreover, DNA double-strand breaks were increased in cancer cells 24 hours post radium-223 treatment in the LuCaP 58 model providing further evidence of anti-tumor effects. Radium-223-treated mice exhibited less visceral metastases in the LuCaP 58 model (not significant). Based on autoradiography, radium-223 was deposited in the intratumoral bone matrix and in conjunction with osteoblasts in osteoblastic metastases. We demonstrate that radium-223 dichloride is successfully incorporated into the intratumoral bone matrix and inhibits tumor growth in both cell line- and patient-derived osteoblastic prostate cancer metastasis models. Given the α-particle range of 50-80 μm, potent radiation effects on the tumor microenvironment are evident whereas relevant effects on the more distant bone marrow are not expected. Taken together, radium-223 therapy exhibits a dual mode-of-action that impacts tumor growth and tumor-induced bone reaction, both important players in the destructive vicious cycle of osteoblastic bone metastasis in prostate cancer. Citation Format: Mari I. Suominen, Katja M. Fagerlund, Jukka P. Rissanen, Yvonne Konkol, Jukka Morko, Zhiqi Peng, Esa Alhoniemi, Dominik Mumberg, Karl Ziegelbauer, Sanna-Maria Käkönen, Jussi M. Halleen, Robert L. Vessella, Arne Scholz. Radium-223 dichloride exhibits dual mode-of-action inhibiting both tumor and tumor-induced bone growth in two osteoblastic prostate cancer models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3447. doi:10.1158/1538-7445.AM2015-3447

Iron (III)-Sodium-EDTA, as used for food fortification, aggravates intestinal inflammation and drives tumorigenesis in mouse models of colitis-associated cancer

Western lifestyle is associated with increasing prevalence of IBD, processed food and food fortification being a likely cause. Iron supplementation may increase gut inflammation and colitis-associated cancer (CAC). Novel Fe products which are chemically not Fe(II) salts and presumably better tolerated have become available. Here we compared various Fe compounds at 450 mg elemental Fe/kg chow [Na-EDTA-Fe(III) (as used for food fortification), Fe(III)maltol (Iron Therapeutics), plant Fe (Biogena), and Fe(II)SO4)], in their potential to promote colitis and CAC in animal models (AOM/DSS and IL-10-/-). In addition, in vitro experiments evaluated ROS production (DCFD assay), DNA damage (yH2AX) and oncogenic pathways in HT29, HCT116, Caco-2, RKO, and 1CT cells. Unexpectedly, Na-EDTA-Fe(III)-treated mice developed the severest form of colitis and only the 1st and last DSS cycles were administered. Similarly, in IL-10-/- mice, the Na-EDTA-Fe(III) group was euthanized 4 months before all other groups due to severe weight loss and disease activity (DAI). The mean DAI in the Na-EDTA-Fe(III) AOM/DSS mice was higher (1.82 vs. 0.94 (control), 0.76 (Fe(II)SO4), 0.67 (Fe(III)maltol), 0.54 (plant Fe), 0.80 (iron deficient); p < 0.001 by ANOVA). The total tumor area was higher with Na-EDTA-Fe(III) (5.96 mm2 vs. 1.38 mm2 (control), 1.88 mm2 (FeSO4), 1.97 mm2 (Fe(III)maltol), 0.98 mm2 (plant Fe), 1.51 mm2 (iron deficient); p = 0.001 by ANOVA) and there were more invasive tumors. ROS production was not elevated but there was an increase in yH2AX with Na-EDTA-Fe(III) and phosphorylation of Akt and Erk. Na-EDTA-Fe(III), as used for fortification in flour and cereals, exacerbates intestinal inflammation and drives tumorigenesis in mouse models of IBD. It is unclear whether this effect is caused by EDTA itself (which also is added to food as Na-EDTA or Na-EDTA-Ca(II)) or only in conjunction with Fe. Nevertheless, our data point to the hazard of food processing and fortification regarding IBD and to the necessity of food safety testing in animal models thereof.

High and low frequency subharmonic imaging of angiogenesis in a murine breast cancer model

This project compared quantifiable measures of tumor vascularity obtained from contrast-enhanced high frequency (HF) and low frequency (LF) subharmonic ultrasound imaging (SHI) to 3 immunohistochemical markers of angiogenesis in a murine breast cancer model (since angiogenesis is an important marker of malignancy and the target of many novel cancer treatments). Nineteen athymic, nude, female rats were implanted with 5×106 breast cancer cells (MDA-MB-231) in the mammary fat pad. The contrast agent Definity (Lantheus Medical Imaging, N Billerica, MA) was injected in a tail vein (dose: 180μl/kg) and LF pulse-inversion SHI was performed with a modified Sonix RP scanner (Analogic Ultrasound, Richmond, BC, Canada) using a L9–4 linear array (transmitting/receiving at 8/4MHz in SHI mode) followed by HF imaging with a Vevo 2100 scanner (Visualsonics, Toronto, ON, Canada) using a MS250 linear array transmitting and receiving at 24MHz. The radiofrequency data was filtered using a 4th order IIR Butterworth bandpass filter (11–13MHz) to isolate the subharmonic signal. After the experiments, specimens were stained for endothelial cells (CD31), vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2). Fractional tumor vascularity was calculated as contrast-enhanced pixels over all tumor pixels for SHI, while the relative area stained over total tumor area was calculated from specimens. Results were compared using linear regression analysis. Out of 19 rats, 16 showed tumor growth (84%) and 11 of them were successfully imaged. HF SHI demonstrated better resolution, but weaker signals than LF SHI (0.06±0.017 vs. 0.39±0.059; p<0.001). The strongest overall correlation in this breast cancer model was between HF SHI and VEGF (r=−0.38; p=0.03). In conclusion, quantifiable measures of tumor neovascularity derived from contrast-enhanced HF SHI appear to be a better method than LF SHI for monitoring angiogenesis in a murine xenograft model of breast cancer (corresponding in particular to the expression of VEGF); albeit based on a limited sample size.

963 Iron (III)-Sodium-EDTA As Used for Food Fortification Aggravates Intestinal Inflammation and Drives Tumorigenesis in Mouse Models of Colitis-Associated Cancer

Background: Western lifestyle is associated with increasing prevalence of IBD, processed food and food fortification being a likely cause. Iron supplementation may increase gut inflammation and colitis-associated cancer (CAC). Novel Fe products which are chemically not Fe(II) salts and presumably better tolerated have become available. Methods: Here we compared various Fe compounds at 450mg elemental Fe/kg chow [Na-EDTA-Fe(III) (as used for food fortification), Fe(III) maltol (Iron Therapeutics Ltd), plant Fe (Biogena, Austria), and Fe(II)SO4)], in their potential to promote colitis and CAC in animal models of IBD (AOM/DSS and IL-10-/-). In addition, experiments were performed in vitro to evaluate their impact on ROS production (DCFD assay), DNA damage ( γH2AX) and activation of oncogenic pathways (in HT29, HCT116, Caco-2 and RKO, and the normal colonic cell line 1CT). Results: Unexpectedly, Na-EDTA-Fe(III)-treated mice developed the severest form of colitis and only the 1st and 4th DSS cycles were administered. Similarly, in IL-10-/mice, the NaEDTA-Fe(III) group was euthanized 4 months before all other groups due to severe weight loss and disease activity (DAI). The mean DAI in the Na-EDTA-Fe(III) AOM/DSS mice was higher (1.82 vs. 0.94 (control), 0.76 (Fe(II)SO4), 0.67 (Fe(III) maltol), 0.54 (plant Fe), 0.80 (ID); p<0.001 by ANOVA). The total tumor area per mouse was higher with Na-EDTAFe(III) (Figure; p=0.001 by ANOVA), and there were more invasive tumors. ROS production was not elevated but there was an increase in γH2AX with Na-EDTA-Fe(III) and phosphorylation of Akt and Erk. Conclusion: Na-EDTA-Fe(III), as it is commonly used for food fortification in flour products and cereals, exacerbates intestinal inflammation and drives tumorigenesis in mouse models of IBD. It is currently unclear whether this toxic effect is caused by EDTA itself (which also is added to food as Na-EDTA or Na-EDTA-Ca(II)) or only in

Sequential intranodal immunotherapy induces antitumor immunity and correlated regression of disseminated follicular lymphoma

AbstractAdvanced stage follicular lymphoma (FL) is incurable by conventional therapies. In the present pilot clinical trial, we explored the efficacy and immunogenicity of a novel in situ immunotherapeutic strategy. Fourteen patients with untreated or relapsed stage III/IV FL were included and received local radiotherapy to solitary lymphoma nodes and intranodal injections of low-dose rituximab (5 mg), immature autologous dendritic cells, and granulocyte-macrophage colony-stimulating factor at the same site. The treatment was repeated 3 times targeting different lymphoma nodes. Primary end points were clinical responses and induction of systemic immunity. Five out of 14 patients (36%) displayed objective clinical responses, including 1 patient with cutaneous FL who showed regression of skin lesions. Two of the patients had durable complete remissions. Notably, the magnitude of vaccination-induced systemic CD8 T-cell–mediated responses correlated closely with reduction in total tumor area (r = 0.71, P = .006), and immune responders showed prolonged time to next treatment. Clinical responders did not have a lower tumor burden than nonresponders pretreatment, suggesting that the T cells could eliminate large tumor masses once immune responses were induced. In conclusion, the combined use of 3 treatment modalities, and in situ administration in single lymphoma nodes, mediated systemic T-cell immunity accompanied by regression of disseminated FL. The trial was registered at www.clinicaltrials.gov as #NCT01926639.

Prognostic factors in renal cancer

A wide variety of histologic prognostic parameters have been proposed for renal cell carcinoma and of these tumor morphotype, tumor grade, sarcomatoid and rhabdoid differentiation, and tumor necrosis were accepted as having utility in clinical practice by the Consensus Conference on Renal Neoplasia assembled by the International Society of Urological Pathology in Vancouver in 2012. With respect to morphotype it was determined that both papillary and chromophobe RCC have a more favourable prognosis than clear cell RCC, while collecting duct RCC was recognised as having a poor prognosis. At the conference it was agreed that for clear cell and papillary RCC grading should based on nucleolar prominence alone for grade 1 to 3 tumours, and that grade 4 tumors should be defined by the presence of extreme nuclear pleomorphism, tumor giant cells and/or rhabdoid/sarcomatoid differentiation. It was further agreed that chromophobe RCC should not be graded. For necrosis there was consensus that the presence or absence of tumour necrosis should be routinely included in the histological report and that this should be based upon both macroscopic and histological examination. There was also consensus that the area of necrosis should be recorded as a percentage of the total tumour area.

Breast cancer size estimation with MRI in BRCA mutation carriers and other high risk patients

ObjectiveTo assess the value of breast MRI in size assessment of breast cancers in high risk patients, including those with a BRCA 1 or 2 mutation. Guidelines recommend invariably breast MRI screening for these patients and therapy is thus based on these findings. However, the accuracy of breast MRI for staging purposes is only tested in sporadic cancers. MethodsWe assessed concordance of radiologic staging using MRI with histopathology in 49 tumors in 46 high risk patients (23 BRCA1, 12 BRCA2 and 11 Non-BRCA patients). The size of the total tumor area (TTA) was compared to pathology. In invasive carcinomas (n=45) the size of the largest focus (LF) was also addressed. ResultsCorrelation of MRI measurements with pathology was 0.862 for TTA and 0.793 for LF. TTA was underestimated in 8(16%), overestimated in 5(10%), and correctly measured in 36(73%) cases. LF was underestimated in 4(9%), overestimated in 5(11%), and correctly measured in 36(80%) cases. Impact of BRCA 1 or 2 mutations on the quality of size estimation was not observed. ConclusionsTumor size estimation using breast MRI in high risk patients is comparable to its performance in sporadic cancers. Therefore, breast MRI can safely be used for treatment planning.

Assessment of Chemotherapy Response in Colorectal Liver Metastases in Patients Undergoing Hepatic Resection and the Correlation to Pathologic Residual Viable Tumor

The Response Evaluation Criteria in Solid Tumors (RECIST), which evaluates maximum tumor diameter only, is commonly used to determine response to chemotherapy in patients with colorectal liver metastases. Limitations of RECIST include its inability to assess the changes in tumor enhancement. The aim of this study was to assess the correlation of these criteria as well as the modified RECIST (mRECIST) with pathologic tumor response. A novel semi-automated volumetric assessment of tumor size was also investigated. A review of a 1,948-patient prospective hepatic database to assess response and pathologic criteria was performed. Patients undergoing preoperative chemotherapy before hepatic resection for colorectal liver metastases were reviewed. Radiographic responses according to RECIST and mRECIST were determined. The percentage of viable tumor cells compared with the total tumor area was determined from the pathologic specimens. We identified 38 patients with adequate imaging who had undergone anatomic hepatic resection and full pathologic evaluation. The percentages of residual viable tumor in the resected specimens were significantly different across RECIST categories (p = 0.045), but not mRECIST (p = 0.305). For mRECIST, there were improved and significant linear trends for residual viable tumor, necrosis, and necrosis + fibrosis when compared with RECIST (p = 0.056). Neither RECIST nor mRECIST responses were predictive of residual viable tumor burden in regression analyses. A novel semi-automated volumetric assessment of tumor size correlated well with pathologic tumor size. Neither RECIST nor mRECIST were predictive of residual viable burden, although the linear trend for mRECIST and residual necrosis + fibrosis compared favorably with RECIST. Continued evaluation for tumor enhancement and standardization of tumor size remain a critical unmet need in patients with solid organ disease.

Effects of Ibuprofen on orthotopic glioma model in rats.

To determine the effects of ibuprofen (Ibp) on the vessel proliferation and necrosis in a rat glioma model. Experimental, randomized interventional trial. 1st Neurosurgery Clinic in Bakirkoy Mental Diseases Hospital, Bakirkoy, Istanbul, Turkey, in the year 2010. After stereotactic injection of C6/LacZ rat glioma cells into the Wistar rats brain, the rats were randomly assigned to two treatment groups (group 1, control; group 2, Ibp treatment). Rats were sacrificed 18 days after treatment, and number of intra-/peri-tumoural vessels, microendothelial proliferations, immunohistochemistry and necrotic area were evaluated. Ibp treatment significantly decreased tumour tissue, intratumoral vessel number and total tumour area level. The level of Ki67 was significantly decreased in the tumour tissue of group 2. Additionally, the total necrotic area / total tumour volume (%) was significantly less in the tumour tissue of the ibuprofen-treated rats compared to the controls. The data show that the Ibp produced an important reduction in glioma tumour cell proliferation in the rat model.

Combination hyperbaric oxygen and temozolomide therapy in c6 rat glioma model

Temozolomide (TMZ) has anti-tumor activity in patients with malignant glioma. Hyperbaric oxygen (HBO) may enhance the efficacy of certain therapies that are limited because of the hypoxic tumor microenvironment. We examined the combined effects of TMZ-HBO in a rat glioma model. After stereotactic injection of C6/LacZ rat glioma cells into the Wistar rats brain, the rats were randomly assigned to three treatment groups [group 1, control treatment; group 2, TMZ alone; group 3, a combination of TMZ and HBO]. Rats were sacrificed 18 days after treatment, and number of intra-/peri-tumoral vessels, microendothelial proliferations, immunohistochemistry and necrotic area were evaluated. Tumoral tissue was stained only sparsely with GFAP. Temozolomide treatment was significantly decreased in tumor tissue intratumoral vessel number / total tumor area level. The level of Ki67 was significantly decreased in the tumor tissue of the group 3. Additionally, the total necrotic area / total tumor volume (%) was decreased significantly in tumor tissue of the group 3 rats compared to group 1 and 2. The combination of hyperbaric oxygen with temozolomide produced an important reduction in glioma growth and effective approach to the treatment of glioblastoma.

Serotonin expression in pancreatic neuroendocrine tumors correlates with a trabecular histologic pattern and large duct involvement

Pancreatic neuroendocrine tumors with prominent stromal fibrosis are often clinically, radiographically, and grossly indistinguishable from ductal adenocarcinoma. We recently described a small series of fibrotic pancreatic neuroendocrine tumors that express serotonin. To understand better the relationship between histopathologic patterns and serotonin expression, we reviewed 361 pancreatic neuroendocrine tumors to identify those with prominent stromal fibrosis exceeding 30% of total tumor area. We identified 52 cases and immunolabeled these neoplasms with antibodies to serotonin and Ki-67. Two predominant histologic subtypes were identified: 14 (26.9%) of 52 had a trabecular or trabecular-glandular cellular pattern with interspersed fibrosis, whereas 38 (73.1%) of 52 had solid architecture. Of the 52, 14 (26.9%) pancreatic neuroendocrine tumors showed at least focal serotonin immunoreactivity. Tumors with predominantly trabecular architecture were significantly more likely to express serotonin than those with solid architecture (P < .01). Only 2 of 34 pancreatic neuroendocrine tumors with fibrosis less than 30% of total tumor area expressed serotonin. The 14 serotonin-expressing tumors were less likely to have lymph node metastases (P = .016) and more likely to involve large pancreatic ducts (P < .01) than were the 38 serotonin-negative tumors. The serotonin-expressing tumors were also found in a younger patient population (P < .01). There was no significant association of serotonin immunoreactivity with Ki-67 proliferation index, tumor size, or distant metastases. Our data demonstrate a strong correlation between trabecular architecture and serotonin immunoreactivity in pancreatic neuroendocrine tumors with stromal fibrosis. Serotonin-expressing tumors are also less likely to have lymph node metastases and more likely to involve large pancreatic ducts.

Abstract B207: Darinaparsin is an anti-solid tumor cytotoxin in vivo.

Abstract Background: Darinaparsin (DPS; provided by ZIOPHARM Oncology, Inc.) is a novel organic arsenical with promising anticancer activity and a favorable systemic toxicity profile. Compared to the inorganic arsenical anti-cancer drug, arsenic trioxide (ATO), DPS has higher in vitro potency against solid tumor cells under both normoxic (NO) and hypoxic (HO) conditions. Under HO, an important feature of the solid tumor microenvironment, DPS activity was not dependent on the generation of reactive oxygen species and subsequent oxidative stress. Here we report results of studies of the in vivo anti-tumor effect of DPS in xenograft solid tumor models and patient-derived prostate cancer tumor grafts, as well as mechanistic studies including cDNA microarray and other experiments to study mechanisms of action of DPS. Materials and Methods: To test the in vivo efficacy of DPS, we used mouse models of human prostatic [hormone-independent (HI) LAPC-4] and pancreatic [PANC-1] xenograft tumors implanted subcutaneously in immunodeficient mice. In addition, patient-derived prostate cancer tissues were precision-cut and implanted as tissue slice grafts (TSGs) under the renal capsule in immunodeficient mice. Both tumor models were treated with DPS (100 mg/kg, IP, 3 times/wk × 4 wks), or saline as a control. The TSG tumor growth was assessed by immunohistochemistry (AMACR/p63) staining, and the results were represented as the total tumor area in TSG sections. The effects of DPS on JNK and HIF1 signaling pathways were determined by western blot and RT-qPCR. Using cDNA microarray and subsequent pathway analyses, we studied the effect of DPS and ATO on global gene expression in HI-LAPC-4 cells. Normal and transformed Mouse Embryonic Fibroblasts (MEFs) overexpressing the oncogene HRAS and E1A were used to study the effect of DPS on these oncogenes that are important for susceptibility to cell killing. Results: In both HI-LAPC-4 and PANC-1 tumor models, DPS significantly inhibited tumor growth (P &amp;lt; 0.0001). The average tumor volume doubling time increased from 3.95 days (95% CI: 4.22 − 3.72) to 11.8 days (95% CI: 11.2 − 12.5) in HI-LAPC-4 tumors (P &amp;lt; 0.001), and 3.83 days (95% CI: 3.65 − 4.02) to 6.82 (95% CI: 6.59 − 7.07) in PANC-1 tumors (P &amp;lt; 0.001). In the TSG model, DPS decreased the tumor growth by 50% in 4 of the 6 TSGs as compared with the paired control (P = 0.139). DPS-treated mice exhibited decreased physical activity and early body weight loss (∼5–10% in the 1st wk), but no other significant systemic toxicities as assessed by organ histology and blood biochemistry. As previously reported in leukemia cells, JNK was activated by DPS under both NO and HO; however, the JNK inhibitor SP600125 did not inhibit DPS activity. HIF1α, a major regulator of cell survival under HO, was not affected by DPS. Microarray analysis revealed a significant effect of DPS (but not ATO) on RAS and MYC oncogenic pathways. Compared to normal MEFs, oncogene (RAS/E1A) transformed MEFs were markedly more sensitive to DPS-induced apoptosis under both NO and HO. Conclusions: DPS had significant cytotoxic activity in vivo in the two tumor models studied (HI-LAPC-4 and PANC-1) in mice at a dose that was well-tolerated. This may be due, in part, to DPS inhibition of mutant and dysregulated oncogenic pathways that are required for survival of tumor, but not normal cells. Since DPS is in early clinical development as a single agent, these findings may have near term translational potential. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr B207.

Spatial relationship of phosphorylated epidermal growth factor receptor and activated AKT in head and neck squamous cell carcinoma

BackgroundOverexpression of EGFR correlates with decreased survival after radiotherapy in head and neck squamous cell carcinoma (HNSCC). However, the contribution of the activated form, pEGFR, and its downstream signaling (PI3-K/AKT) pathway is not clear yet. MethodsFifty-eight patients with HNSCC were included in the study. pEGFR, pAKT, hypoxia, and vessels were visualized using immunohistochemistry. Fractions (defined as the tumor area positive for the respective markers relative to the total tumor area) were calculated by automated image analysis and related to clinical outcome. ResultsBoth pEGFR (median 0.6%, range 0–34%) and pAKT (median 1.8%, range 0–16%) expression differed between tumors. Also, a large variation in hypoxia was found (median pimonidazole fraction 3.9% 0–20%). A significant correlation between pEGFR and pAKT (rs 0.44, p=0.004) was seen, however, analysis revealed that this was not always based on spatial coexpression. Low pAKT expression was associated with increased risk of regional recurrence (p<0.05, log-rank) and distant metastasis (p=0.04). ConclusionThe correlation between expression of pEGFR and pAKT is indicative of activation of the PI3-K/AKT pathway through phosphorylation of EGFR. Since not all tumors show coexpression to the same extent, other factors must be involved in the activation of this pathway as well.

Initial analytic validation of automated bone scan measures for treatment response assessment in patients with metastatic castration-resistant prostate cancer (CRPC).

e15174 Background: Evaluation of novel therapies for patients with CRPC has been limited by the lack of validated quantitative measures of bone tumor burden. Tools including visual assessment for determining progression based on lesion count and semi quantitative measures including the Bone Scan Index for predicting survival have been proposed. To date, a validated quantitative measure that can be used to assess either progression or response has not been available. We have developed a computer-aided (CAD) system for measurement of bone scan tumor burden including lesion count (LC), mean intensity (MI) and total area (TA). This study is to assess the range of normal variation and potential clinically meaningful cut points for changes in bone scan tumor burden. Methods: From an anonymized image data set, 20 CRPC patients were selected with a pair of bone scans acquired 6 to 24 weeks apart. All patients had failed at least 2nd line drug therapy; 10 were on no therapy, 10 received drug therapy between the two scans. Scans were acquired at 12 sites using a standard of care protocol including 20-25 mCi of Tc MDP, injected 2-4 hours before whole body scintigraphy. An automated CAD system was run for each patient on bone scans. Degenerative joint disease was removed manually by one of two board certified Nuclear Medicine Physicians. A two-sample Wilcoxon rank sum (Mann-Whitney) test was used to test for differences between the two groups in the ratio of LC (visit 2 / visit 1), change in MI and % change in tumor TA. Results: Table shows the median (IQR) for ratio of LC, change in MI and % change in TA. Differences between groups were significant for all measures. Conclusions: This preliminary data suggests total bone scan tumor area may be useful as an early surrogate outcome for CRPC patients with 30% change from baseline as a potential cut point to distinguish responders from non-responders. This work lays a foundation for validation of these measures against other clinically relevant outcome measures in a larger cohort. Ratio of LC Change in MI % change in TA No therapy 1.24 (0.56) -1.40 (14.80) 7.13 (27.61) Drug therapy 0.54 (0.68) -17.60 (16.30) -73.76 (45.38) P value 0.0012 0.0019 0.0003

Pathologic Response of Primary Rectal Cancer to Oxaliplatin-Based Chemotherapy

Management of stage IV rectal cancer is controversial, and different strategies may be useful. Preoperative chemotherapy for liver metastases might cause pathologic changes over the primary rectal tumor. In this study, the authors show the pathologic regression of the primary rectal tumor after neoadjuvant chemotherapy treatment. Patients suffering stage IV rectal cancer underwent surgery after oxaliplatin-based chemotherapy. Age, gender, type of surgery, carcinoembryogenic antigen (CEA) level, presence of metastatic disease in one or multiple organs, ypT, ypN, and circumferential resection margin (CRM) were evaluated. Pathologic response of the primary tumor was estimated by using three conventional grading systems and a semiquantitative system assessed by the amount of viable cells out of the total tumor area macroscopically described. Fibrosis, necrosis, and colloid response were evaluated with a semiquantitative system. A complete pathologic response (ypTO) was found in one patient. A good response was observed in the 41.6% of the cases with all grading systems. Presence of fibrosis in the primary tumor was found in six cases. No patient showed CRM involvement. One patient developed a local recurrence. Oxaliplatin-based chemotherapy for stage IV rectal cancer provides high rates of pathologic regression in the rectal tumor and may allow surgery without CRM involvement.