Squamous cell carcinoma (SCC) can be treated by using different strategies, including photodynamic therapy (PDT).PDT is a non-invasive therapy that consists in the combination of visible light, molecular oxygen and a photoactive compound (photosensitizer) that leads to the formation of reactive oxygen species, inducing cell death.Tumoral microenvironment (TME) has gained prominence over the last few years since it has been indicated to be crucial for tumor progression as well as for the response to treatments.One of the TME components are cancer-associated fibroblasts (CAFs).CAFs are activated fibroblasts characterized by the

OBJECTIVES: Recent evidence has suggested that sub-classifying endocervical adenocarcinomas based on histopathologic characteristics, including degree of destructive stromal invasion and desmoplastic response, may help further refine early risk stratification. We sought to explore the translational prognostic implications of the above histopathologic stratification in women undergoing treatment for early stage cervical adenocarcinoma.

Abstract The human breast tissue consists of lobules connected to a complex network of ducts that are evolutionarily designed to produce and transport milk to nourish offspring. Histopathology has identified 10 major cell types based on morphological features but have provided limited information on cell states - the transcriptional programs of cell types that reflect different biological functions. In this study, we have generated an unbiased 'cell atlas' of the normal human breast to define the cell types and cell states using single cell RNA sequencing methods. We performed 3' microdroplet based single cell RNA sequencing of 31,442 stromal cells from 11 women with pathologically normal breast tissues that were collected from mastectomies. Unbiased expression analysis identified three major cell types: epithelial cells (luminal and basal), fibroblasts and endothelial cells, in addition to several minor cell types: macrophages, T-cells, natural killer cells, pericytes and smooth muscle cells. Analysis of cell states of these cell types revealed different transcriptional programs in luminal epithelial cells (hormone receptor positive and secretory), basal epithelial cells (myoepithelial or basement-like), endothelial cells (lymphatic or vascular), macrophages (M1 or M2) and fibroblasts (three subgroups) and provided insight into progenitors of each cell types. These data provide a valuable reference for the research community and will provide new insights into how normal cell types are transformed in the tumor microenvironment to promote or inhibit the progression of breast cancer. Citation Format: Seth TK, Bai S, Hu M, Sei E, Wood A, Wiley J, Chen H, Contreras A, Teshome M, Lim B, Navin NE. Towards a human breast cell atlas [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr GS1-02.

Analysis of somatic mutations in solid tumors and hematologic malignancies using targeted next generation sequencing (NGS)-based assays has become part of routine oncology practice as well as clinical trials. The use of paired tumor-normal DNA samples increases confidence of somatic calls. NGS assays that utilize unique patient identifiers (SNP IDs) allow further comparison of samples within a run or paired tumor/normal samples. The sources of germline DNA include peripheral blood (PB) and formalin-fixed paraffin-embedded tissue (FFPE). However, the source of normal can be problematic, especially in transplant setting. Herein, we report two cases of NGS-based molecular testing in a patient with mycosis fungoides treated with stem cell transplant [SCT] (Pt1) and a patient with lung adenocarcinoma who previously had acute leukemia cured by SCT. These cases highlight the importance of selecting an appropriate normal sample for excluding germline polymorphisms during somatic mutation testing. Initial analyses that included concurrent PB sample failed to filter known germline polymorphisms. Repeat analyses using pre-transplant PB/bone marrow allowed for the successful subtraction of germline variants. Somatic mutations in PTEN and ERBB4 (Pt1) and CDKN2A, KRAS, KDR, and TP53 (Pt2) were reported with confidence. Selection of an appropriate source of germline DNA for NGS-based somatic mutation testing for patients with SCT transplant can be challenging. Particular attention to the clinical history is crucial for accurate interpretation and reporting.

Deregulation and activation of the phosphoinositide 3-kinase (PI3K)/Akt/mammalian (or mechanistic) target of rapamycin (mTOR) pathway have a major role in proliferation and cell survival in breast cancer. However, as single agents, mTOR inhibitors have had modest antitumor efficacy. In this study, we evaluated the effects of vertical inhibition of mTOR and Akt in breast cancer cell lines and xenografts. We assessed the effects of mTOR inhibitor rapamycin and Akt inhibitor MK-2206, given as single drugs or in combination, on cell signaling, cell proliferation and apoptosis in a panel of cancer cell lines in vitro. The antitumor efficacy was tested in vivo. We demonstrated that MK-2206 inhibited Akt phosphorylation, cell proliferation and apoptosis in a dose-dependent manner in breast cancer cell lines. Rapamycin inhibited S6 phosphorylation and cell proliferation, and resulted in lower levels of apoptosis induction. Furthermore, the combination treatment inhibited phosphorylation of Akt and S6, synergistically inhibited proliferation and induced apoptosis with a higher efficacy. In vivo combination inhibited tumor growth more than either agent alone. Our data suggest that a combination of Akt and mTOR inhibitors have greater antitumor activity in breast cancer cells, which may be a viable approach to treat patients.

C laboratories are rapidly implementing next-generation sequencing (NGS) tests for mutation analysis, however there are few guidelines regarding sample quality for successful results. We aimed to establish tissue quality parameters for successful NGS in solid tumors and to improve NGS performance. Using a 50-gene hotspot mutation panel we identified the major cause for unsuccessful NGS analysis being DNA 10 mm2. Independent factors leading to lower NGS success were cellular tumor areas and decalcification procedures. Tumor type and paraffin block age did not affect success. We optimized workflow and showed improved NGS success rates with pronounced improvement among tiny samples and cytology samples. Identifying preanalytical tissue factors allows us to improve NGS performance and to successfully test tumors obtained from minimally invasive procedures.

Radiation resistance is a critical factor in treatment failure after lung cancer therapy. We evaluated protein factors that could confer radiation resistance phenotype in lung cancer cells and validated these proteins for clinical outcomes in patients who had postoperative radiation therapy after complete resection. Clonogenic Survival Assay (CSA) was performed on a panel (60) of non-small cell lung cancer cell lines (NSCLC) and reverse phase protein array (RPPA) was used to determine protein markers that correlated best with survival fraction after 2 Gy (SF2). Tissue Microarray (TMA) from NSCLC surgical specimen from 128 patients who underwent postoperative radiation therapy for locally advanced disease was performed for the most predictive proteins and quantified using a 4-value intensity score and percentage of the extent of reactivity. Univariable and multivariable (MVA) time to event analysis were performed to associate median TMA staining levels to locoregional failure survival (LRFS), distant metastatic–free survival (DMFS), and overall survival (OS). High SF2 correlated with higher levels of Ku80, BRCA1, ATR, UPF1, RAD50, pATR, pSrc, pACC, and lower levels of Bax, pTSC2, pAKT, and pMAPK. Some of these DNA damage repair (DDR) proteins were coordinately expressed with UPF1, a protein involved in nonsense-mediated decay (NMD) of mRNAs but also has been reported to interact with DDR proteins. RNAi knockdown (KD) of UPF1 caused significant downregulation of XRCC1, Chk1, RAD50 and ERCC1 in A549 and H1299 NSCLC cell lines. UPF1 KD, and not UPF2 KD, also sensitized H1299 cell line to radiation. High UPF1 nuclear staining in tumors trended for worse LRF (P=.067), but high nuclear RAD50 significantly correlated with worse LRF (P=.007), DMFS (P=.008) and OS (P=.036). On MVA, UPF1 is no longer predictive, but high RAD50 levels (vs low by median) remains significant for LRF (HR 1.62, 95% CI 1.02-2.55), DMFS (HR 1.60, 95% CI 1.02-2.52), but not OS (HR 1.46, 95% CI 0.92-2.31). Greater age and number of positive pN2 nodes were also independent predictors of LRF, DMFS, and OS. DDR proteins correlate with radiation resistance in NSCLC cell lines, and UPF1 KD downregulates these DDR genes in concert. While UPF1 levels in tumors do not itself correlate with disease outcomes in patients, downregulating its levels may reduce RAD50 whose expression is prognostic in patients. UPF1 could serve as an attractive target to enhance radiation sensitization in NSCLC.