Nitrilase 1 modulates lung tumor progression in vitro and in vivo.

Uncovering novel growth modulators for non-small cell lung cancer (NSCLC) may lead to new therapies for these patients. Previous studies suggest Nit1 suppresses chemically induced carcinogenesis of the foregut in a mouse model. In this study we aimed to determine the role of Nit1 in a transgenic mouse lung cancer model driven by a G12D Kras mutation. Nit1 knockout mice (Nit1-/-) were crossed with KrasG12D/+ mice to investigate whether a G12D Kras mutation and Nit1 inactivation interact to promote or inhibit the development of NSCLC. We found that lung tumorigenesis was suppressed in the Nit1-null background (Nit1-/-:KrasG12D/+). Micro-CT scans and gross tumor measurements demonstrated a 5-fold reduction in total tumor volumes compared to Nit1+/+KrasG12D/+ (

Targeting fibroblast activation protein in tumor stroma with chimeric antigen receptor T cells can inhibit tumor growth and augment host immunity without severe toxicity.

The majority of chimeric antigen receptor (CAR) T-cell research has focused on attacking cancer cells. Here, we show that targeting the tumor-promoting, nontransformed stromal cells using CAR T cells may offer several advantages. We developed a retroviral CAR construct specific for the mouse fibroblast activation protein (FAP), comprising a single-chain Fv FAP [monoclonal antibody (mAb) 73.3] with the CD8α hinge and transmembrane regions, and the human CD3ζ and 4-1BB activation domains. The transduced muFAP-CAR mouse T cells secreted IFN-γ and killed FAP-expressing 3T3 target cells specifically. Adoptively transferred 73.3-FAP-CAR mouse T cells selectively reduced FAP(hi) stromal cells and inhibited the growth of multiple types of subcutaneously transplanted tumors in wild-type, but not FAP-null immune-competent syngeneic mice. The antitumor effects could be augmented by multiple injections of the CAR T cells, by using CAR T cells with a deficiency in diacylglycerol kinase, or by combination with a vaccine. A major mechanism of action of the muFAP-CAR T cells was the augmentation of the endogenous CD8(+) T-cell antitumor responses. Off-tumor toxicity in our models was minimal following muFAP-CAR T-cell therapy. In summary, inhibiting tumor growth by targeting tumor stroma with adoptively transferred CAR T cells directed to FAP can be safe and effective, suggesting that further clinical development of anti-human FAP-CAR is warranted

Non Small Cell Carcinoma of the Lung- From Morphology to Molecular Profiling

Dr. Charalambos Solomides is an Associate Professor of Pathology at the Thomas Jefferson University Hospital. At present, he holds many positions at Jefferson. He is a Staff Surgical Pathologist, Staff Cytopathologist, Director of Immunnohistochemistry Research Lab, Director of Cytopathology and the Director of Fine Needle Aspiration Services. His interests are in Non-small cell lung carcinoma, radiation induced lung injury and idiopathic pulmonary fibrosis

Correlations between Gene Amplification and Protein Expression of Topoisomerase 2A (TOP2A) in Squamous Cell Carcinoma of the Lung

Background: While DNA topoisomerase 2A (TOP2A) plays an essential role in maintaining the structural integrity of the double helix during replication and recombination, excessive expression of this enzyme may promote malignant cell transformations. In fact, increased levels of TOP2A have been observed in various cancer cell lines including squamous cell carcinoma of the lung. This study sought to identify correlations between genotypic and phenotypic evidence of TOP2A obtained via in situ hybridization (ISH) and immunohistochemistry (IHC) techniques. Methods Tissue microarrays created from 29 samples of Stage I Squamous Cell Carcinoma of the lung were stained with VENTANA BenchMark ULTRA platform with dual color ISH molecular probes TOP2A / CEP17 and antiTOP2A antibody (clone JS5B4-rabbit monoclonal antibody). Gene copy numbers were analyzed using bright field microscopy. Gene amplification was considered in cells exhibiting gene copies \u3e 3 or TOP2A:CEP17 ratios \u3e 1.82. IHC stains were quantified using Spectrum software (Apeiro technologies) using the nuclear algorithm. All levels of protein expression (+1 to +3) were considered positive. Results: A moderate Pearson Correlation (0.4) between TOP2A gene amplification and protein expression was identified. Conclusion: While gene amplification moderately correlated with protein expression of TOP2A, additional factors influencing protein expression independently of gene amplification should be identified

Comparison of Two Quantitative Image Analysis Systems for Breast Cancer Immunohistochemistry

Automated image analysis systems for breast cancer immunohistochemistry promise efficiency and reliability in the quantification of therapy targets such as the estrogen receptor (ER) or human epidermal growth factor receptor (Her2). Thomas Jefferson University Hospital owns two such systems, the Aperio ScanScope AT (Leica Biosystems) and Ventana iScan Coreo (Roche). A comparison study was performed to determine if choice of system affects target quantification and subsequent clinical tumor classification. Tumor expressions of ER, progesterone receptor (PR), proliferation marker Ki67, and Her2 were quantified with both systems for tissue samples from twenty breast cancer patients. Positive tumor classification was based on percent positivity values of ER \u3e1%, PR \u3e1%, Ki67 \u3e10%, and Her2 score of 3+. Percent agreement for tumor classification was ER 100 (95% CI 83.2-100), PR 95 (75.1-99.9), Ki67 90 (68.3-98.8) and Her2 100 (83.2-100). While agreement was high large variation was observed in percent positivity scores (ER, PR, Ki67)

Diagnostic Value of Endoscopic Ultrasound-Guided Fine Needle Aspiration of Intra-Abdominal Lymph Nodes in Patients with Concurrent Biopsy of Intra-Abdominal Tumors

BACKGROUND • Diagnostic endoscopic biopsy of intra-abdominal lesions may be performed prior to surgical resections. • Endoscopic ultrasound-guided fine needle aspiration of intra-abdominal lymph nodes may also be performed to: • Yield more information • Allow for more accurate staging • We evaluate the additional staging information concurrent fine needle aspiration of intra-abdominal lymph nodes provided DESIGN • We included all patients at our institution from January 1, 2000 to March 30, 2015 who, during the same endoscopic procedure, had: • Endoscopic ultrasound guided fine needle aspiration of an intra-abdominal lymph node AND • Endoscopic ultrasound guided fine needle aspiration OR a surgical biopsy of an intraabdominal lesion • We excluded all patients for whom the final diagnosis was lympho-proliferative • Primary lesions were: • Pancreatic • Upper gastrointestinal tract • Biliary tract • Gallbladder • 63 total patient

Cytohistologic correlation of basaloid salivary gland neoplasms: Can cytomorphologic classification be used to diagnose and grade these tumors?

BACKGROUND: Basaloid salivary gland neoplasms (BSNs), which include benign primary tumors and primary or metastatic malignancies, show overlapping morphology in fine-needle aspiration (FNA). The Milan system recommends assigning a grade (low or high) to malignant salivary neoplasms because of the impact on surgical planning. This study investigated cytomorphologic features of BSNs on FNA that would help to favor a high-grade malignancy over a low-grade malignancy or a benign tumor. METHODS: Two pathologists performed a double-blinded cytologic evaluation of FNA cases diagnosed as BSNs that had corresponding surgical resections. The diagnosis made with the Milan system was correlated with the final surgical diagnosis and grade. Cytologic sensitivity, specificity, and predictive values were calculated. RESULTS: There were 132 BSN FNA cases; cytology slides were available for 77 of 87 patients who had undergone resection. The risk of malignancy for the benign neoplasm (BN), salivary gland neoplasm of uncertain malignant potential (SUMP), suspicious for malignancy (SFM), and malignant categories were 13.6%, 22%, 100%, and 100%, respectively. The sensitivity of the malignant/SFM category was 51.7%; another 37.9% of confirmed malignancies were diagnosed as SUMP. The specificity of the BN category was 86%. Favoring a high-grade malignancy on FNA had 100% accuracy (5 of 5). Favoring a low-grade malignancy on FNA had 75% accuracy (6 of 8). The most specific cytomorphologic clues for a high-grade malignancy were necrotic/apoptotic debris, mitoses, discohesion, and anisonucleosis. CONCLUSIONS: BSNs encompass a broad spectrum of primary and metastatic tumors. Necrotic/apoptotic debris, mitotic activity, discohesion, and significant anisonucleosis, alone or especially in combination, should make a cytopathologist suspect a high-grade malignancy