CD44+/EPCAM+ cells detect a subpopulation of ALDHhigh cells in human non-small cell lung cancer: A chance for targeting cancer stem cells?

Objectives: Several studies demonstrated that aldehyde dehydrogenase (ALDH) and CD44 are the most considered cancer stem cells (CSC) markers. However, a comparison between ALDH high cells and CD44+ cells have been previously described with no significant correlation. Indeed, the aim of the present research is to identify a superficial marker able to match with ALDH high cells population in freshly isolated human lung cancer cells. Materials and Methods: This cross-sectional study analyzed the expression of ALDHhigh/low cells and the positivity for CD44 and epithelium cell adhesion molecule (EPCAM) antigens in surgical lung cancer tissues. The main approach was a cytofluorimetric analysis of ALDH expression and positivity for CD44/EPCAM on primary cell population obtained from 23 patients harboring NSCLC. Results: There was a highly positive correlation between the expressions of ALDHhigh and CD44+/EPCAM+ cells, with a Pearson\u2019s correlation coefficient equal to 0.69 (95% CI 0.39\u20130.86; P = 0.0002), and Spearman\u2019s correlation coefficient equal to 0.52 (P = 0.0124). The average paired difference between the expression of ALDHhigh and CD44+/EPCAM+ cells was very close to 0, being 0.1% (SD 2.5%); there was no difference between these subpopulations in terms of means (95% CI = \u20131.0; 1.2%, P = 0.8464). These results highlight a strong similarity between ALDHhigh and CD44+/EPCAM+ cells. Conclusions: Our study is the first attempt which identifies a high correlation between the ALDHhigh and the CD44+/EPCAM+ cells, thus suggesting the possibility to use this superficial marker for future target treatments against lung cancer stem cells.Objectives: Several studies demonstrated that aldehyde dehydrogenase (ALDH) and CD44 are the most considered cancer stem cells (CSC) markers. However, a comparison between ALDH high cells and CD44+ cells have been previously described with no significant correlation. Indeed, the aim of the present research is to identify a superficial marker able to match with ALDH high cells population in freshly isolated human lung cancer cells. Materials and Methods: This cross-sectional study analyzed the expression of ALDHhigh/low cells and the positivity for CD44 and epithelium cell adhesion molecule (EPCAM) antigens in surgical lung cancer tissues. The main approach was a cytofluorimetric analysis of ALDH expression and positivity for CD44/EPCAM on primary cell population obtained from 23 patients harboring NSCLC. Results: There was a highly positive correlation between the expressions of ALDHhigh and CD44+/EPCAM+ cells, with a Pearson\u2019s correlation coefficient equal to 0.69 (95% CI 0.39\u20130.86; P = 0.0002), and Spearman\u2019s correlation coefficient equal to 0.52 (P = 0.0124). The average paired difference between the expression of ALDHhigh and CD44+/EPCAM+ cells was very close to 0, being 0.1% (SD 2.5%); there was no difference between these subpopulations in terms of means (95% CI = \u20131.0; 1.2%, P = 0.8464). These results highlight a strong similarity between ALDHhigh and CD44+/ EPCAM+ cells. Conclusions: Our study is the first attempt which identifies a high correlation between the ALDHhigh and the CD44+/EPCAM+ cells, thus suggesting the possibility to use this superficial marker for future target treatments against lung cancer stem cells

Isolation and Identification of Cancer Stem-Like Cells in Adenocarcinoma and Squamous Cell Carcinoma of the Lung: A Pilot Study

Background: Lung cancer stem cells (CSCs) share many characteristics with normal stem cells, such as self-renewal and multipotentiality. High expression of aldehyde dehydrogenase (ALDH) has been detected in many tumors, particularly in the CSC compartment, and it plays an important role in tumor proliferation, metastasis, and drug resistance. CD44 is commonly used as a cell surface marker of cancer stem-like cells in epithelial tumors. The aim of this study was to isolate and analyze cancer stem-like cells from surgically removed specimens to compare lung adenocarcinoma (ADENO) and squamous (SQUAMO) cell carcinoma. Methods: The ALDEFLUOR assay was used to identify and sort ALDHhigh and ALDHlow human lung cancer cells following tissue digestion. Fluorescence-activated cell sorting analysis for CD44 was performed with tumor cells. Quantitative real-time PCR was performed to assess the expression of SOX2 and NANOG as stemness markers. ALDH1A1 expression was additionally determined by immunohistochemistry. Anchorage-independent ALDHhigh cell growth was also evaluated. ALDHhigh ADENO and SQUAMO cells were cultured to analyze spheroid formation. Results: All specimens contained 0.5–12.5% ALDHhigh cells with 3.8–18.9% CD44-positive cells. SOX2 and NANOG relative expression in ALDHhigh compared to ALDHlow cells in ADENO and SQUAMO was analyzed and compared between the histotypes. Immunohistochemistry confirmed the presence of ALDH1A1 in the sections. SOX2 and NANOG were expressed at higher levels in the ALDHhigh subpopulation than in the ALDHlow subpopulation only in ADENO cells, and the opposite result was seen in SQUAMO cells. In vitro functional assays demonstrated that ALDHhigh cells exhibited migration capacity with distinct behaviors between ALDHhigh spheres in ADENO vs. SQUAMO samples. Conclusions: Our results highlight the importance of a better characterization of cancer stem-like cells in ADENO and SQUAMO histotypes. This may suggest new differential approaches for prognostic and therapeutic purposes in patients with non-small-cell lung cancer

Cancer Stem-Like Cells in a Case of an Inflammatory Myofibroblastic Tumor of the Lung.

Background: Inflammatory myofibroblast tumor (IMT) is a rare tumor with obscure etiopathogenesis in which different inflammatory cells and myofibroblastic spindle cells are seen histologically. Although the majority of these neoplasms have a benign clinical course, the malignant form has also been reported. The gold standard is surgical treatment for complete removal. Our report describes a 50-year-old woman who underwent surgery for IMT of the lung. The aim is to determine whether cancer stem cells may be present in IMT of the lung. Methods: In April 2018, the patient underwent surgery for tumor mass asportation through lateral thoracotomy. The histology of the tumor was consistent with IMT of the lung. The ALDEFLUOR assay, after tissue digestion, was used to identify and sort human lung cancer cells expressing high and low aldehyde dehydrogenase (ALDH) activity. SOX2, NANOG, OCT-4, and c-MYC positivity were additionally determined by immunohistochemistry. Results: The specimen contained 1.10% ALDHhigh cells among all viable lung cancer cells, which indicates the population of cancer stem cells is not negligible. Immunohistochemically assessed cell positivity for ALDH1A1, SOX2, NANOG, OCT-4, and c-MYC, which are considered as lung cancer stem-like cells markers. Conclusion: For the first time, we demonstrated the presence of cancer stem cells in a case of IMT of the lung. This finding may provide a base for considering new pathological and molecular aspects of this tumor. This perspective suggests further studies to understand the possibility of developing recurrence depending on the presence of cancer stem cells

Enhancing Lightpath QoT Computation with Machine Learning in Partially Disaggregated Optical Networks

Increasing traffic demands are causing network operators to adopt disaggregated and open networking solutions to better exploit optical transmission capacity, and consequently enable a software-defined networking (SDN) approach to control and management that encompasses the WDM data transport layer. In these frameworks, a quality of transmission estimator (QoT-E) that gives the generalized signal-to-noise ratio (GSNR) is commonly used to compute the feasibility of transparent lightpaths (LP)s, taking into account the amplified spontaneous emission (ASE) noise and the nonlinear interference (NLI). In general, the ASE noise is the main contributor to the GSNR and is also the most challenging noise component to evaluate in a scenario with varying spectral loads, due to fluctuations in the optical amplifier responses. In this work, we propose a machine learning (ML) algorithm that is trained using different ASE-shaped spectral loads in order to predict the OSNR component of the GSNR; this methodology is subsequently used in combination with a QoT-E in the lightpath computation engine (L-PCE). We present an experiment on a point-to-point optical line system (OLS), including 9 commercial erbium-doped fiber amplifiers (EDFA)s used as black-boxes, each with variable gain and tilt values, and 8 fibers that are characterized by distinct physical parameters. Within this experiment, we receive the signal at the end of the OLS, measuring the bit-error-rate (BER) and the power spectrum, over 2520 different spectral loads. From this dataset, we extract the expected GSNRs and their linear and nonlinear components. Through joint application of a ML algorithm and the open-source GNPy library, we obtain a complete QoT-E, demonstrating that a reliable and accurate LP feasibility predictor may be implemented

Experimental Demonstration of Partially Disaggregated Optical Network Control Using the Physical Layer Digital Twin

Optical communications and networking are fast becoming the solution to support ever-increasing data traffic across all segments of the network, expanding from core/metro networks to 5G/6G front-hauling. Therefore, optical networks need to evolve towards an efficient exploitation of the infrastructure by overcoming the closed and aggregated paradigm, to enable apparatus sharing together with the slicing and separation of the optical data plane from the optical control. In addition to the advantages in terms of efficiency and cost reduction, this evolution will increase network reliability, also allowing for a fine trade-off between robustness and maximum capacity exploitation. In this work, an optical network architecture is presented based on the physical layer digital twin of the optical transport used within a multi-layer hierarchical control operated by an intent-based network operating system. An experimental proof of concept is performed on a three-node network including up to 1000 km optical transmission, open re-configurable optical add & drop multiplexers (ROADMs) and whitebox transponders hosting pluggable multirate transceivers. The proposed solution is based on GNPy as the optical physical layer digital twin and ONOS as intent-based network operating system. The reliability of the optical control decoupled by the data plane functioning is experimentally demonstrated exploiting GNPy as open lightpath computation engine and software optical amplifier models derived from the component characterization. Besides the lightpath deployment exploiting the modulation format evaluation given a generic traffic request, the architecture reliability is tested mimicking the use case of an automatic failure recovery from a fiber cut

Comparison of HER-2 and hormone receptor expression in primary breast cancers and asynchronous paired metastases: impact on patient management

The assessment of hormone receptors (HRs) and human epidermal growth factor receptor (HER)-2 is necessary to select patients who are candidates for hormonal and anti-HER-2 therapy. The evaluation of these parameters is generally carried out in primary tumors and it is not clear if reassessment in metastatic lesions might have an impact on patient management. The primary aim of this analysis was to compare HER-2 and HR status in primary tumors versus metastatic sites in breast cancer patients. PATIENTS AND METHODS: Seventy-five patients with available samples from primary tumors and paired metastases were included. HER-2 status was evaluated by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH); HR status was assessed by IHC. RESULTS: Nineteen percent of primary tumors were HER-2 positive; 77% were HR positive. Sites of biopsied or resected metastases were: locoregional soft tissues (n = 30), liver (n = 20), central nervous system (n = 5), bone (n = 5), pleura (n = 4), distant soft tissues (n = 3), abdomen (stomach, colon, peritoneum) (n = 3), bronchus (n = 3), and bone marrow (n = 2). For paired metastases, the HER-2 status was unchanged in 84% of cases; two patients changed from positive to negative, while 10 patients converted from negative to positive (agreement, 84%; kappa = 0.5681). A change in HR status was observed in 16 cases (21%): nine cases from positive to negative and seven cases from negative to positive (agreement, 78.7%; kappa = 0.4158). CONCLUSIONS: Further studies are necessary to better define the level of discordance in HER-2 or HR status between primary tumors and paired metastases. However, a biopsy of metastatic disease can be recommended, if feasible with minimal invasiveness, because treatment options might change for a significant proportion of patient

Comprehensive analysis of Moringa oleifera leaves’ antioxidant properties in ovine cheese

This study aimed to enhance “Pecorino” type ovine cheese by adding Moringa oleifera leaves powder (MOLP). Cheese-making trials, conducted at industrial level, used raw ewes’ milk and two selected Lactococcus lactis strains. The experimental plan included a control production (CTR), and two experimental productions with 1% or 2% MOLP addition (1-MOLP and 2-MOLP, respectively). MOLP did not hinder starters development, which reached about 8.0 Log CFU/g in 2-month ripened cheeses. Illumina results highlighted lactococci dominance in all trials [45.98%–62.48% of relative abundance (RA)]. Physicochemical analysis showed that MOLP-enriched cheeses had higher protein content and lower secondary lipid oxidation. The addition of MOLP increased total phenolic compounds in cheese, reaching 3.64 mg GAE/g in the 2-MOLP sample. MOLP-enriched cheeses showed significantly higher radical scavenging activity than CTR production (p < 0.0001). Ultrahigh-performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) revealed increased levels of chlorogenic acid, protocatechuic acid, caffeic acid, and ferulic acid due to MOLP enrichment. In the presence of MOLP, cheese volatile organic compounds were affected by compounds like 2-octanone, 3-hexen-2-one, heptane, nonanol, and linalool. 1-MOLP cheese was comparable to CTR production in overall satisfaction (sensory evaluation). Including MOLP in cheese production offers exciting opportunities for functional Sicilian ewes’ milk products

PIK3CA Mutation in the ShortHER Randomized Adjuvant Trial for Patients with Early HER2\ufe Breast Cancer: Association with Prognosis and Integration with PAM50 Subtype.

Purpose: We explored the prognostic effect of PIK3CA mutation in HER2\ufe patients enrolled in the ShortHER trial. Patients and Methods: The ShortHER trial randomized 1,253 patients with HER2\ufe breast cancer to 9 weeks or 1 year of adjuvant trastuzumab combined with chemotherapy. PIK3CA hotspot mutations in exon 9 and 20 were analyzed by pyrosequencing. Expression of 60 genes, including PAM50 genes was measured using the nCounter platform. Results: A mutation of the PIK3CA gene was detected in 21.7% of the 803 genotyped tumors. At a median follow-up of 7.7 years, 5-year disease-free survival (DFS) rates were 90.6% for PIK3CA mutated and 86.2% for PIK3CA wild-type tumors [HR, 0.84; 95% confidence interval (CI), 0.56\u20131.27; P \ubc 0.417]. PIK3CA mutation showed a favorable prognostic impact in the PAM50 HER2-enriched subtype (n \ubc 232): 5-year DFS 91.8% versus 76.1% (log-rank P \ubc 0.049; HR, 0.46; 95% CI, 0.21\u20131.02). HER2-enriched/PIK3CA mutated versus wild-type tumors showed numerically higher tumor-infiltrating lymphocytes (TIL) and significant upregulation of immune-related genes (including CD8A, CD274, PDCD1, and MYBL2, a proliferation gene involved in immune processes). High TILs as well as the upregulation of PDCD1 and MYBL2 were associated with a significant DFS improvement within the HER2-enriched subtype (HR, 0.82; 95% CI, 0.68\u20130.99; P \ubc 0.039 for 10% TILs increment; HR, 0.81; 95% CI, 0.65\u20130.99; P \ubc 0.049 for PDCD1 expression; HR, 0.72; 95% CI, 0.53\u20130.99; P \ubc 0.042 for MYBL2 expression). Conclusions: PIK3CA mutation showed no prognostic impact in the ShortHER trial. Within the HER2-enriched molecular subtype, patients with PIK3CA mutated tumors showed better DFS versus PIK3CA wild-type, which may be partly explained by upregulation of immune-related genes