Tumor Quantification Research Articles

Prognostic utility of epidermal growth factor receptor (EGFR) expression in prostatic acinar adenocarcinoma

BackgroundEpidermal growth factor receptor (EGFR) is potential prognostic biomarker expressed in many human cancers. Prognostic significance of EGFR immunohistochemical expression has not been established in prostatic acinar adenocarcinoma, therefore we aimed to evaluate the frequency of expression of EGFR in prostatic adenocarcinoma and its association with other prognostic parameters.MethodsThe study included 123 cases of biopsy proven prostatic acinar adenocarcinoma treated at Liaquat National hospital, Karachi from January 2013 till December 2017. Paraffin blocks of all cases were retrieved; sections were cut and stained with haematoxylin and eosin. Pathologic characteristics including tumor quantification, WHO grade group, gleason score, perineural and lymphovascular invasion were evaluated. EGFR immunohistochemistry (IHC) was performed on all tissue blocks.ResultsMean age of the patients included in the study was 69.05 ± 8.68 years. High gleason scores i.e. 8 & 9 were noted in 22% (27 cases) and 22.8% (28 cases) respectively. Similarly, 22.8% (28 cases) showed WHO grade group 5. 52.8% (65 cases) had > 50% tissue involvement by carcinoma and perineural invasion was seen in 37.4% (46 cases). Positive EGFR expression was noted in 18.7% (23 cases), while 81.3% (100 cases) showed negative EGFR expression. Significant association of EGFR expression was noted with gleason score (p-value = < 0.001), WHO grade (p = < 0.001), tumor quantification (p = 0.007) and perineural invasion (p = < 0.001). Moreover, significant association of EGFR expression was also seen with disease recurrence and Her2neu over expression. Patients with low gleason scores (score 6 and 7) and lower grade group (1, 2 & 3) were less likely to have positive EGFR expression as compared to patients with high gleason score (score 9) and higher grade group (5). Similarly, patients with perineural invasion were more likely to have positive EGFR expression.ConclusionWe found a relatively low EGFR expression in our patients with prostatic adenocarcinoma; however, its association with poor prognostic parameters like high gleason score, higher grade group, perineural invasion, higher tissue involvement by cancer and disease recurrence signifies its importance as a prognostic parameter in prostatic acinar adenocarcinoma.

Intraoperative visualization of the tumor microenvironment and quantification of extracellular vesicles by label-free nonlinear imaging.

Characterization of the tumor microenvironment, including extracellular vesicles (EVs), is important for understanding cancer progression. EV studies have traditionally been performed on dissociated cells, lacking spatial information. Since the distribution of EVs in the tumor microenvironment is associated with cellular function, there is a strong need for visualizing EVs in freshly resected tissues. We intraoperatively imaged untreated human breast tissues using a custom nonlinear imaging system. Label-free optical contrasts of the tissue, correlated with histological findings, enabled point-of-procedure characterization of the tumor microenvironment. EV densities from 29 patients with breast cancer were found to increase with higher histologic grade and shorter tumor-to-margin distance and were significantly higher than those from 7 cancer-free patients undergoing breast reduction surgery. Acquisition and interpretation of these intraoperative images not only provide real-time visualization of the tumor microenvironment but also offer the potential to use EVs as a label-free biomarker for cancer diagnosis and prognosis.

Respiratory motion correction in prostate cancer positron emission tomography: A study on patients and phantom simulation

Introduction: To investigate the effects of breathing cycle and tree diaphragm motions on prostate cancer tumors standard uptake value (SUV) during positron emission tomography (PET) and to correct it. Materials and methods: Respiratory motion traces were simulated on the common patient breathing cycle and tree diaphragm motions using the 4D NCAT phantom. Lesions with different activities and diameters were simulated in prostate. This study was conducted in two phases. In the first phase; the generated PET sinograms taken from GATE (Geant4 Application Tomography Emission) Monte Carlo simulations corrected using computed tomography attenuation correction involving the cycle. In the next phase; on the obtained images, the motion-correction was carried out using software for Tomographic Image Reconstruction (STIR). By considering respiration- averaged computed as a true value, the SUVs were measured and analyzed for different motions. Results: In uncorrected measurements, the mean, minimum and maximum SUVs were 0.4%, 0.09% and 2% respectively, whilst in corrected measurements, these value were reported as 3.5%, 0.7% and 14% respectively. Conclusion: The respiratory motion-induced errors in tumor quantification and delineation. The SUVs depending on the phase of the raspatory. Motion correction could enhance the SUVs in prostate cancer PET.

Histopathologic quantification of viable tumor versus treatment effect in surgically resected recurrent glioblastoma.

The prognostic impact of the histopathologic features of recurrent glioblastoma surgical specimens is unknown. We sought to determine whether key histopathologic characteristics in glioblastoma tumors resected after chemoradiotherapy are associated with overall survival (OS). The following characteristics were quantified in recurrent glioblastoma specimens at our institution: extent of viable tumor (accounting for % of specimen comprised of tumor and tumor cellularity), mitoses per 10 high-power fields (0, 1-10, > 10), Ki-67 proliferative index (0-100%), hyalinization (0-6; none to extensive), rarefaction (0-6), hemosiderin (0-6), and % of specimen comprised of geographic necrosis (0-100%; converted to 0-6 scale). Variables associated with OS in univariate analysis, as well as age, eastern cooperative oncology group performance status (ECOG PS), extent of repeat resection, time from initial diagnosis to repeat surgery, and O6-methylguanine-DNA methyltransferase promoter methylation, were included in a multivariable Cox proportional hazards model. 37 specimens were assessed. In a multivariate model, high Ki-67 proliferative index was the only histopathologic characteristic associated with worse OS following repeat surgery for glioblastoma (hazard ratio (HR) 1.3, 95% CI 1.1-1.5, p = 0.003). Shorter time interval from initial diagnosis to repeat surgery (HR 1.11, 95% CI 1.02-1.21, p = 0.016) and ECOG PS ≥ 2 (HR 4.19, 95% CI 1.72-10.21, p = 0.002) were also independently associated with inferior OS. In patients with glioblastoma undergoing repeat resection following chemoradiotherapy, high Ki-67 index in the recurrent specimen, short time to recurrence, and poor PS are independently associated with worse OS. Histopathologic quantification of viable tumor versus therapy-related changes has limited prognostic influence.

Abstract 1594: A fast and sensitive workflow to screen therapy-relevant mutations in circulating tumor cells and quantification of cancer-associated extracellular vesicle-derived mRNA in plasma of cancer patients

Abstract Liquid biopsy can be considered a test that detects rare cells or nucleic acids circulating in blood or in other biofluids. The concept of liquid biopsy aims to monitor the status of a disease or treatment efficacy in a simple, fast, cost efficient way and at any point in time with acceptable risk and burden for the patient. However, the volumes of available blood samples can be quite limited and rare cells as well as circulating nucleic acids are normally present in low abundance. Therefore, detection of rare events or the quantification of limited material requires robust, highly sensitive technologies like droplet digital PCR (ddPCR). Here, a novel workflow is presented combining the effective enrichment of rare cells using the Parsortix microfluidic filtration system with the analysis by ddPCR. Ten or 100 cells of NSCLC cell line NCI-H441 were spiked into a suspension of 100k NCI-H1563 cells or healthy EDTA blood. The resulting cell suspensions were processed using the microfluidic device Parsortix to enrich cell factions by size and deformability. Enriched cell fractions were harvested or lysed directly in the Parsortix cassette. Genomic DNA was prepared from all cell fractions. NCI-H441 cells are heterozygous for KRAS G12V and TP53 R158L mutations, while NCI-H1563 cells do not contain the mutations. Ten cells or less were enough to detect KRAS G12V or TP53 R158L mutations in a standard blood sample (7-10 ml). This combined workflow enables quantification of circulating tumor cells and analysis for therapy relevant point mutations in a fast, EpCAM independent manner. Additionally, extracellular vesicle derived mRNA (EVmRNA) isolated from either cell culture supernatant or plasma from cancer patients was analyzed to demonstrate the suitability of ddPCR for assessing specific EVmRNA as biomarkers in different cancer types. In 13 NSCLC and 17 metastatic breast cancer plasma samples the levels of epithelial and oncogene EVmRNA, including EpCAM, ERBB2 or FGFR3 were determined and compared to healthy control plasma. Levels of potentially cancer-associated EVmRNA were only detectable in plasma from cancer patients, but not in healthy controls. Samples from patients and healthy volunteers, respectively, were collected under signed informed consent. We demonstrated, that ddPCR allows the verification of therapy relevant point mutations in CTCs has the potential to be used as a screening approach. Moreover, despite the small amounts of mRNA in extracellular vesicles from patient blood, using ddPCR differential mRNA levels can be determined in patient plasma and enable and expand the scope of biomarker analysis from EVs. This work is supported by IMI JU &amp; EFPIA (grand no. 115749). Citation Format: Martin H. Neumann, Sebastian Bender, Thomas Krahn, Thomas Schlange. A fast and sensitive workflow to screen therapy-relevant mutations in circulating tumor cells and quantification of cancer-associated extracellular vesicle-derived mRNA in plasma of cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1594.

Early postoperative delineation of residual tumor after low-grade glioma resection by probabilistic quantification of diffusion-weighted imaging.

In WHO grade II low-grade gliomas (LGGs), early postoperative MRI (epMRI) may overestimate residual tumor on FLAIR sequences. Consequently, MRI at 3-6 months follow-up (fuMRI) is used for delineation of residual tumor. This study sought to evaluate if integration of apparent diffusion coefficient (ADC) maps permits an accurate estimation of residual tumor early on epMRI. From a consecutive cohort, 43 cases with an initial surgery for an LGG, and complete epMRI (< 72 hours after resection) and fuMRI including ADC maps, were retrospectively identified. Residual FLAIR hyperintense tumor was manually segmented on epMRI and corresponding ADC maps were coregistered. Using an expectation maximization algorithm, residual tumor segments were probabilistically clustered into areas of residual tumor, ischemia, or normal white matter (NWM) by fitting a mixture model of superimposed Gaussian curves to the ADC histogram. Tumor volumes from epMRI, clustering, and fuMRI were statistically compared and agreement analysis was performed. Mean FLAIR hyperintensity suggesting residual tumor was significantly larger on epMRI compared to fuMRI (19.4 ± 16.5 ml vs 8.4 ± 10.2 ml, p < 0.0001). Probabilistic clustering of corresponding ADC histograms on epMRI identified subsegments that were interpreted as mean residual tumor (7.6 ± 10.2 ml), ischemia (8.1 ± 5.9 ml), and NWM (3.7 ± 4.9 ml). Therefore, mean tumor quantification error between epMRI and fuMRI was significantly reduced (11.0 ± 10.6 ml vs -0.8 ± 3.7 ml, p < 0.0001). Mean clustered tumor volumes on epMRI were no longer significantly different from the fuMRI reference (7.6 ± 10.2 ml vs 8.4 ± 10.2 ml, p = 0.16). Correlation (Pearson r = 0.96, p < 0.0001), concordance correlation coefficient (0.89, 95% confidence interval 0.83), and Bland-Altman analysis suggested strong agreement between both measures after clustering. Probabilistic segmentation of ADC maps facilitates accurate assessment of residual tumor within 72 hours after LGG resection. Multiparametric image analysis detected FLAIR signal alterations attributable to surgical trauma, which led to overestimation of residual LGG on epMRI compared to fuMRI. The prognostic value and clinical impact of this method has to be evaluated in larger case series in the future.

Abstract 2024: Hyaluronic acid conjugated nanoparticle delivery of siTWIST reduces tumor burden and enhances chemosensitivity in ovarian cancer

Abstract Purpose: TWIST is a transcription factor critical to embryonic development, and aberrantly activated in many cancers. It regulates epithelial-to-mesenchymal transition (EMT), the process underlying metastatic spread and drug resistance. The majority of epithelial ovarian cancer (EOC) patients respond well to chemotherapy, but relapse with metastatic and drug-resistant disease. We are investigating the role of TWIST-mediated relapse, with the goal of developing treatments that sensitize chemoresistant tumors and improve survival. Experimental Procedures: We employ siRNA to target TWIST mRNA and have created a mesoporous silica nanoparticle with hyaluronic acid (siTWIST-MSN-HA) as a delivery vehicle. Microscopy was conducted to ensure cancer stem cell (CSC) targeting via CD44, and efficient uptake of MSN-HAs. We tested TWIST knockdown combined with cisplatin in vitro and in vivo. At necropsy, total tumor, metastasis, and ascites were evaluated to determine effects of siTWIST-MSN-HA compared to cisplatin alone. qPCR was conducted on tumors to examine effect of siRNA against TWIST, and its target genes. Mice were imaged via bioluminescence to observe CSC localization of siTWIST-MSN-HA compared to siTWIST-MSN (No HA). Tissue sections were stained via IHC to determine MSN-HA tumor targeting. CD44 staining was conducted to reveal HA co-localization in tumor. Results: siTWIST-MSN-HAs significantly knocked down TWIST, and sensitized cells to cisplatin compared to control in vitro. Following necropsy, tumor quantification revealed mice treated with siTWIST-MSN-HAs plus cisplatin exhibited a startling 75% loss of overall tumor burden (p=0.0012), 88% loss of total metastasis (p=0.001), and further 86% loss of total ascites (p=0.002) compared to cisplatin-alone treatment groups. EMT target expression by qPCR analysis revealed loss of TWIST1, vimentin, N-cadherin, and gain of E-cadherin in tumors treated with siTWIST-MSN-HA compared to controls. Bioluminescent images of mice at necropsy revealed highly specific tumor localization of MSN-HA Dylight (680nm) compared to MSN without HA. MSN-HA exhibited CSC targeting at metastatic sites, where most of the signal was emitted from the primary tumor; and negligible quantities of MSN-HAs were detected elsewhere. MSN-HA RITC (576nm) reveals highly specific tumor targeting via IHC; MSN-HA nanoparticles localized primarily in tumor cells and not in control organs. CD44 staining of these tissues reveals MSN-HAs co-localized with CD44 in CSCs. Conclusions: These studies demonstrate TWIST as a promising target for metastasis and acquired drug resistance in EOC. MSN-HAs provide CSC specific CD44 targeting with high efficacy and reduce tumor burden via siTWIST therapy. Thus, siTWIST-MSN-HAs provide a promising platform to improve survival from metastatic, drug-resistant ovarian cancer. Citation Format: Carlotta A. Glackin, Sophia A. Shahin, Shirleen Simargi, Ruining Wang, Altagracia Contreras, Liliana Parra, Loiuse Qu, Wei Wen, Thanh Dellinger, Julia Unternaehrer, Fuyujiko Tamanoi, Jeffrey Zink. Hyaluronic acid conjugated nanoparticle delivery of siTWIST reduces tumor burden and enhances chemosensitivity in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2024.

Drug quantification in metastatic brain tumors in patients receiving bendamustine as a radiosensitizer for stereotactic radiotherapy.

e14017Background: In this Phase I study, we evaluated the ability of the DNA alkylating agent, bendamustine (BD), to serve as a radiosensitizer in patients receiving stereotactic radiotherapy (SRT)...

In vivo near infrared fluorescence imaging and dynamic quantification of pancreatic metastatic tumors using folic acid conjugated biodegradable mesoporous silica nanoparticles

Cancer metastasis is one of the biggest challenges in cancer treatments since it increases the likelihood that a patient will die from the disease. Therefore, the availability of techniques for the early detection and quantification of tumors is very important. We have prepared cyanine 7.5 NHS ester (Cy7.5) and folic acid (FA) conjugated biodegradable mesoporous silica nanoparticles (bMSN@Cy7.5-FA NPs) (~100 nm) for visualizing tumors in vivo. The fluorescence spectra revealed that the emission peak of bMSN@Cy7.5-FA NPs had a red-shift of 1 nm. Confocal immunofluorescent images showed that bMSN@Cy7.5-FA NPs had an excellent targeting ability for visualizing cancer cells. In vivo fluorescence imaging has been conducted using an orthotopic model for pancreatic cancer within 48 h, and the fluorescence intensity reached a maximum at a post injection time-point of 12 h, which demonstrated that the use of bMSN@Cy7.5-FA NPs provides an excellent imaging platform for tumor precision therapy in mice.

Nuclei Segmentation for Quantification of Brain Tumors in Digital Pathology Images

In this article, based on image transformation of HSV (Hue, Saturation, Value), the authors propose a method for cancer nuclei segmentation when such conflicts of cancer nuclei involve ‘omics' indicative of brain tumors pathologically. To constrain the problem space in the region of color information, i.e. cancer nuclei, they convert the images into the V component of HSV first, and then apply the threshold level-set segmentation and the sparsity technique (VTLS-ST) in segmentation. The combined technique of the proposed VTLS-ST is implemented using the real-time CBTC dataset in the validation stage. The proposed method exhibits an improved capability of searching recursively for the optimal threshold level-set in the working subsets via the sparsity representation in segmentation. The experimental results show the reliability and efficiency of the proposed approach in real-time applications with an average rate of 0.932 in terms of similarity index for segmentation of cancer nuclei in brain tumor detection.

MRI-Based Radiation Therapy: Intrafraction Motion Quantification of Head and Neck Tumors Using Cine Magnetic Resonance Imaging

MRI-Based Radiation Therapy: Intrafraction Motion Quantification of Head and Neck Tumors Using Cine Magnetic Resonance Imaging

Whole body PD-L1 PET in patients with NSCLC and melanoma.

139 Background: PD-(L)1 immunotherapy is effective in multiple tumors, including NSCLC and melanoma, but tumor PD-L1 IHC correlates only moderately with treatment outcome. This study aims to assess 1) safety of 18F-BMS-986192 (18F-PD-L1) in human, 2) PD-L1 quantification in tumors using 18F-PD-L1 PET, 3) PD-L1 PET correlation with IHC and treatment outcome, and 4) intra and inter subject tracer uptake variability. Methods: Pts with NSCLC (N = 10) and melanoma (N = 3) were included. At baseline, pts received a static or multiphase dynamic whole body PET scan after injecting 200 MBq 18F-BMS-986192. For NSCLC pts, (1) SUV(max, peak and mean) were measured for each delineable tumor (N = 32, 1-7 tumors/pt), (2) PD-L1 IHC (28.8 assay) was performed on the biopsy, and (3) response to Nivolumab therapy assessed by RECIST 1.1. Intra and inter subject variability and intraclass correlation were calculated using SUVs of all assessed tumors. Equal variance for PD-L1 status was evaluated by a Levene’s test. Four (3 female) pts underwent dosimetry study (ICRP 60). Results: No AEs related to radiotracer was observed. Dosimetry study demonstrated whole body exposure of 30 mGy at dose &gt; 1400 MBq. Biodistribution among pts is comparable. PD-L1 IHC from 13 biopsied lesions were evaluated, 5 &lt; 1%, 4 ≥1%, and 4 ≥50%. Tumor tracer uptake was measured in NSCLC pts and categorized by PDL-1 IHC as ≥50% or &lt; 50%. Clinical trial information: 2015-004760-11. Tumor SUVs did not correlate with RECIST 1.1 assessment. Lesion heterogeneity was reflected in both inter and intra pt variability (CVinter = 41%, CVintra = 53%, ICC = 0.41 for SUVpeak). Levene’s test showed no significance in variability between the two PD-L1 categories. Conclusions: PET-imaging with 18F-BMS-986192 is safe and feasible in pts with NSCLC and melanoma. Pts with higher PD-L1 PET SUV have higher PD-L1 by IHC. Intra pt variability is similar to inter pt variability. With limited number of pts, no clear correlation of PET PD-L1 and tumor response is observed. A prospective study with this tracer is underway to further investigate 18F-BMS-986192 in understanding of PD-L1 expression.[Table: see text]

Abstract B171: PK/PD modeling of the first-in-class, potent and selective covalent CDK7 inhibitor, SY-1365, provides mechanistic basis for intermittent dosing regimens in preclinical efficacy models of hematologic and solid tumors

Abstract Introduction: CDK7 has recently emerged as an attractive gene control target in cancers driven by transcriptional dependencies, including triple-negative breast cancer (TNBC) and acute myeloid leukemia (AML). SY-1365 is a first-in-class, potent, and selective covalent CDK7 inhibitor that has shown durable and complete responses in xenograft models of AML and various solid tumors. To advance the development of this CDK7 inhibitor in patients, the current work assessed the relationship between SY-1365 pharmacokinetics (PK), pharmacodynamics (PD) measured as CDK7 target occupancy, and efficacy in mouse xenograft models of AML (HL-60) and TNBC (HCC70). Methods: SY-1365 was dosed intravenously in several xenograft experiments across a range of dose levels (5-40 mg/kg) and dose frequencies (daily to weekly). PK and changes in CDK7 occupancy were measured following single and repeat dosing, and efficacy studies measured the resultant dose-dependent changes in tumor volume over time after repeat dosing of SY-1365. To account for the temporal differences in PK, PD and tumor growth inhibition (TGI), a mathematical modeling approach was taken to integrate the resulting datasets. In addition, an assay to determine CDK7 occupancy in human PBMCs was developed to support the ongoing phase 1 study. Results: The PK in mouse was proportional with respect to dose and exhibited a terminal elimination half-life (t1/2) of 4 h. The PK/PD model was able to describe the PK, PD, and efficacy in HL-60 and HCC70 xenografts, with similar exposure and CDK7 occupancy profiles observed in these two xenografts. The model demonstrated a CDK7 turnover t1/2 of 69 h, longer than that reported for other oncogenic kinases targeted with covalent inhibitors where daily dosing is required (e.g., EGFRi, BTKi), with optimal efficacy achieved when target engagement was sustained over the dosing interval. This provides a mechanistic basis for intermittent dosing based on the dose-dependent TGI, including tumor regressions, observed with SY-1365 in in vivo models of TNBC and AML. Conclusions: The PK/PD relationship for target engagement and efficacy of the first-in-class potent and selective covalent CDK7 inhibitor, SY-1365, has been characterized in preclinical models through a mathematical modeling approach. Assays were developed to enable target occupancy quantification in xenograft tumors, and in human PBMCs as a surrogate tissue to support clinical investigation. Further investigations into tumor molecular profiles driving response are currently under way. In summary, this work supported the selection of an initial twice-weekly dosing regimen in the ongoing phase 1 trial of SY-1365 in adult patients with advanced solid tumors (NCT03134638). Citation Format: Nigel J. Waters, Shanhu Hu, Brett Matzuka, Graeme Hodgson, Yixuan Ren, Yoon Choi, Kevin Dykstra, Christopher Roberts, Kevin Sprott, Emmanuelle di Tomaso, Christian Fritz. PK/PD modeling of the first-in-class, potent and selective covalent CDK7 inhibitor, SY-1365, provides mechanistic basis for intermittent dosing regimens in preclinical efficacy models of hematologic and solid tumors [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr B171.

Prognostic Value of FBXO39 and ETS-1 but not BMI-1 in Iranian Colorectal Cancer Patients

Background:Colorectal cancer (CRC) is one of the most prevalent cancers worldwide. Despite recent progress in diagnosis and treatment, it remains a major health problem and further studies are needed. We here investigated expression profiles of the FBXO39, ETS-1 and BMI-1 genes in CRCs to validate any possible diagnostic/prognostic significance.Material and Methods:Thirty six patients with locally advanced CRC admitted to Hazrate-Rasoul Hospital-Tehran were enrolled. Initially the expression pattern of FBXO39, ETS-1 and BMI-1 genes were determined using RT-PCR in CRC tumor and adjacent normal tissues then real-time RT-PCR was employed to quantify BMI-1 gene expression.Results:FBXO39 expression was restricted to tumor tissues. Interestingly, expression of this gene was detected in all stage-0 tumor samples. There was a significant relation between FBXO39 gene expression and lymph node involvement. The ETS-1 gene was expressed in 66% of all tumor tissues with p-value=0.03 for increase as compared to the adjacent normal samples. In addition, there was a significant relation between ETS-1 gene expression and tumor size and lymph node involvement. RT-PCR demonstrated BMI-1 gene expression in both tumor and normal tissues and quantification by real-time RT-PCR showed no association between BMI-1 levels and CRC clinicopathological features.Conclusion:Expression of FBXO39 and ETS-1 with lymph node involvement may be considered as an alarm for the occurrence of CRC metastasis, and therfore have prognostic value while BMI-1 appears without importance.

Prostate cancer reporting and staging: needle biopsy and radical prostatectomy specimens

Prostatic adenocarcinoma remains the most common cancer affecting men. A substantial majority of patients have the diagnosis made on thin needle biopsies, most often in the absence of a palpable abnormality. Treatment choices ranging from surveillance to radical prostatectomy or radiation therapy are largely driven by the pathologic findings in the biopsy specimen. The first part of this review focuses on important morphologic parameters in needle biopsy specimens that are not covered in the accompanying articles. This includes tumor quantification as well as other parameters such a extraprostatic extension, seminal vesicle invasion, perineural invasion, and lymphovascular invasion. For those men who undergo radical prostatectomy, pathologic stage and other parameters are critical in prognostication and in determining the appropriateness of adjuvant therapy. Staging parameters, including extraprostatic extension, seminal vesicle invasion, and lymph node status are discussed here. Surgical margin status is also an important parameter and definitions and reporting of this feature are detailed. Throughout the article the current reporting guidelines published by the College of American Pathologists and the International Collaboration on Cancer Reporting are highlighted.

Does PET Reconstruction Method Affect Deauville Score in Lymphoma Patients?

When evaluating 18F-FDG PET images with the Deauville score (DS), the quantification of tumor and reference organs limits the problem of optical misinterpretation. Compared with conventional reconstruction algorithms, point-spread function (PSF) modeling increases SUVs significantly in tumors but only moderately in the liver, which could affect the DS. We investigated whether the choice of the reconstruction algorithm affects the DS and whether discordance affects the capability of 18F-FDG PET to stratify lymphoma patients. Methods: Overall, 126 patients with diffuse large B-cell lymphoma were included (56 female and 70 male; median age, 65 y; range, 20-88 y). PET data were reconstructed with the unfiltered PSF method. Additionally, a 6-mm filter was applied to PSF images to meet the requirements of the EANM Research Ltd. (EARL) harmonization program from the European Association of Nuclear Medicine (EANM) (PSFEARL). One hundred interim PET (i-PET) and 95 end-of-treatment PET (EoT-PET) studies were analyzed. SUVmax in the liver and aorta was determined using automatic volumes of interest and compared with SUVmax in the residual mass with the highest 18F-FDG uptake. Results: For i-PET, using PSF and PSFEARL, we classified patients as responders and nonresponders in 60 and 40 cases versus 63 and 37 cases, respectively. Five cases of major discordance (5.0%) occurred (i.e., changes from responder to nonresponder). For Eot-PET, patients were classified using PSF and PSFEARL as responders and nonresponders in 69 and 26 cases versus 72 and 23 cases, respectively. Three cases of major discordance (3.2%) occurred. Concordance (Cohen unweighted κ) between the PSF and the PSFEARL DS was 0.82 (95% confidence interval, 0.73-0.91) for i-PET and 0.89 (95% confidence interval, 0.81-0.96) for EoT-PET. The median follow-up periods were 28.4 and 27.4 mo for i-PET and EoT-PET, respectively. Kaplan-Meier analysis showed statistically significant differences in progression-free survival and overall survival among responders and nonresponders no matter which reconstruction was used for i-PET and EoT-PET. Conclusion: Neither DS nor risk stratification of diffuse large B-cell lymphoma patients is affected by the choice of PET reconstruction. Specifically, the use of PSF is not an issue in routine clinical processes or in multicenter trials. These findings have to be confirmed in escalation and deescalation procedures based on early i-PET.

Assessment of Oncological Outcomes After Radical Prostatectomy According to Preoperative and Postoperative Cancer of the Prostate Risk Assessment Scores: Results from a Large, Two-center Experience

BackgroundAmong prostate cancer (PCa) patients undergoing radical prostatectomy (RP) and with virtually identical unfavorable pathological characteristics, those deemed at low risk (LR) preoperatively had better oncological outcomes than those with intermediate (IR) or high risk (HR) preoperatively. ObjectiveTo examine if this phenomenon still applies when preoperative Cancer of the Prostate Risk Assessment (CAPRA) scores are compared to postoperative scores (CAPRA-S) in RP patients. Design, setting, and participantsWe evaluated 10 290 PCa patients who underwent RP at tertiary care centers in Hamburg (Germany) and Milan (Italy) during 1991–2011. Outcome measurements and statistical analysisAll patients were classified as CAPRA/CAPRA-S LR, IR, or HR (≤2, 3–5, and ≥6 points, respectively). Kaplan-Meier and Cox regression analyses were performed to assess the risk of biochemical recurrence (BCR) and metastatic disease (metD). Results and limitationsOverall, 59.6%, 29.3%, and 11.1% patients were CAPRA-S LR, IR, and HR respectively. For CAPRA-S LR patients, 5-yr BCR-free and metD-free rates for preoperative CAPRA LR versus IR/HR patients were 93.1% versus 85.7% (p<0.001) and 99.7% versus 98.9% (p=0.017), respectively. For CAPRA-S IR/HR patients, the corresponding rates were 69.9% versus 57.2% and 98.2 versus 93.7% (both p<0.001). On multivariable Cox regression analyses, a combination of CAPRA-S and preoperative CAPRA resulted in increases in predictive accuracy for BCR (from 76.0% to 78.3%) and metD (from 82.0% to 84.0%). Specifically, biopsy Gleason patterns and the percentage of positive cores added information to the CAPRA-S score. Long-term follow-up is needed to discern clinical metD differences between preoperative CAPRA risk groups. ConclusionsCAPRA-S LR patients in the CAPRA IR/HR group had higher BCR/metD risk. Conversely, CAPRA-S IR/HR patients in the CAPRA LR group had lower BCR/metD risk. Future tools should incorporate better tumor and Gleason quantification to optimize prediction. Patient summaryWe demonstrated that among 10290 European patients with prostate cancer who underwent radical prostatectomy, those deemed at low risk preoperatively had better oncological outcomes than their peers with intermediate or high risk, despite virtually identical unfavorable pathological characteristics.

Notch3 as a novel therapeutic target in metastatic medullary thyroid cancer

Medullary thyroid cancer portends poor survival once liver metastasis occurs. We hypothesize that Notch3 overexpression in medullary thyroid cancer liver metastasis will decrease proliferation and growth of the tumor. TT cells were modified genetically to overexpress Notch3 in the presence of doxycycline, creating the TT-Notch3 cell line. Mice were injected intrasplenically with either TT-Notch3 or control vector TT-TRE cells. Each cell line had 3 treatment groups: control with 12 weeks of standard chow, early DOX with doxycycline chow at day 0 and for 70 days thereafter, and late DOX with doxycycline chow at 8 weeks. Each animal underwent micro-computed tomography to evaluate for tumor formation and tumor quantification was performed. Animals were killed at 12 weeks, and the harvested liver was stained with Ki-67, hematoxylin and eosin, and Notch3. Induction of Notch3 did not prevent formation of medullary thyroid cancer liver metastases as all mice in the early DOX group developed tumors. However, induction of Notch after medullary thyroid cancer liver tumor formation decreased tumor size, as seen on micro-computed tomography scans (late DOX group). This translated to a 37-fold decrease in tumor volume (P = .001). Notch3 overexpression also resulted in decreased Ki-67 index (P = .038). Moreover, Notch3 induction led to increased areas of neutrophil infiltration and necrosis on hematoxylin and eosin staining of the tumors CONCLUSION: Notch3 overexpression demonstrates an antiproliferative effect on established metastatic medullary thyroid cancer liver tumors and is a potential therapeutic target in treatment.

Computer-assisted stereology and automated image analysis for quantification of tumor infiltrating lymphocytes in colon cancer

BackgroundPrecise prognostic and predictive variables allowing improved post-operative treatment stratification are missing in patients treated for stage II colon cancer (CC). Investigation of tumor infiltrating lymphocytes (TILs) may be rewarding, but the lack of a standardized analytic technique is a major concern. Manual stereological counting is considered the gold standard, but digital pathology with image analysis is preferred due to time efficiency. The purpose of this study was to compare manual stereological estimates of TILs with automatic counts obtained by image analysis, and at the same time investigate the heterogeneity of TILs.MethodsFrom 43 patients treated for stage II CC in 2002 three paraffin embedded, tumor containing tissue blocks were selected one of them representing the deepest invasive tumor front. Serial sections from each of the 129 blocks were immunohistochemically stained for CD3 and CD8, and the slides were scanned.Stereological estimates of the numerical density and area fraction of TILs were obtained using the computer-assisted newCAST stereology system. For the image analysis approach an app-based algorithm was developed using Visiopharm Integrator System software. For both methods the tumor areas of interest (invasive front and central area) were manually delineated by the observer.ResultsBased on all sections, the Spearman’s correlation coefficients for density estimates varied from 0.9457 to 0.9638 (p < 0.0001), whereas the coefficients for area fraction estimates ranged from 0.9400 to 0.9603 (P < 0.0001). Regarding heterogeneity, intra-class correlation coefficients (ICC) for CD3+ TILs varied from 0.615 to 0.746 in the central area, and from 0.686 to 0.746 in the invasive area. ICC for CD8+ TILs varied from 0.724 to 0.775 in the central area, and from 0.746 to 0.765 in the invasive area.ConclusionsExact objective and time efficient estimates of numerical densities and area fractions of CD3+ and CD8+ TILs in stage II colon cancer can be obtained by image analysis and are highly correlated to the corresponding estimates obtained by the gold standard based on stereology. Since the intra-tumoral heterogeneity was low, this method may be recommended for quantifying TILs in only one histological section representing the deepest invasive tumor front.

Low calcium diet increases 4T1 mammary tumor carcinoma cell burden and bone pathology in mice.

Breast cancer metastasizes to bone in the majority of patients with advanced disease. We investigated the effects of inadequate dietary calcium (Ca) on bone turnover, tumor growth, and bone response to tumor in tibia inoculated with 4T1 mammary carcinoma cells. Nine-month-old female Balb/c mice were placed on an adequate Ca (5 g/kg diet, n = 30) or low Ca (80 mg/kg diet, n = 31) diet for 14 days, then injected intratibially with 1,000 4T1 cells (transfected with luciferase for bioluminescence imaging), and sacrificed at 5, 10, or 21 days post-inoculation (n = 7–10 mice/group). Control mice (n = 6/group) were injected with carrier and sacrificed at 10 days post-inoculation. Tibiae with muscle intact were excised and evaluated by microcomputed tomography and histology. In vivo bioluminescent imaging revealed that 4T1 cells metastasized to lung. Therefore, lungs were removed for quantification of tumor. Mice fed low Ca exhibited higher bone turnover and higher tibial lesion scores than mice fed adequate Ca. Lesion severity, manifested as cortical osteolysis and periosteal woven bone formation, and tumor cell infiltration to muscle, increased with time, irrespective of diet. However, for most skeletal endpoints the rates of increase were greater in mice consuming low Ca compared to mice consuming adequate Ca. Infiltration of tumor cells into adjacent muscle, but not metastasis to lung, was also greater in mice consuming low Ca diet. The findings suggest that high bone turnover due to Ca insufficiency results in greater local mammary tumor cell growth, cortical osteolysis, woven bone formation, and invasion to muscle in mice.