Transformation Of Cancer Cells Research Articles

Genetic Ablation of MiR-22 Fosters Diet-Induced Obesity and NAFLD Development.

miR-22 is one of the most abundant miRNAs in the liver and alterations of its hepatic expression have been associated with the development of hepatic steatosis and insulin resistance, as well as cancer. However, the pathophysiological roles of miR-22-3p in the deregulated hepatic metabolism with obesity and cancer remains poorly characterized. Herein, we observed that alterations of hepatic miR-22-3p expression with non-alcoholic fatty liver disease (NAFLD) in the context of obesity are not consistent in various human cohorts and animal models in contrast to the well-characterized miR-22-3p downregulation observed in hepatic cancers. To unravel the role of miR-22 in obesity-associated NAFLD, we generated constitutive Mir22 knockout (miR-22KO) mice, which were subsequently rendered obese by feeding with fat-enriched diet. Functional NAFLD- and obesity-associated metabolic parameters were then analyzed. Insights about the role of miR-22 in NAFLD associated with obesity were further obtained through an unbiased proteomic analysis of miR-22KO livers from obese mice. Metabolic processes governed by miR-22 were finally investigated in hepatic transformed cancer cells. Deletion of Mir22 was asymptomatic when mice were bred under standard conditions, except for an onset of glucose intolerance. However, when challenged with a high fat-containing diet, Mir22 deficiency dramatically exacerbated fat mass gain, hepatomegaly, and liver steatosis in mice. Analyses of explanted white adipose tissue revealed increased lipid synthesis, whereas mass spectrometry analysis of the liver proteome indicated that Mir22 deletion promotes hepatic upregulation of key enzymes in glycolysis and lipid uptake. Surprisingly, expression of miR-22-3p in Huh7 hepatic cancer cells triggers, in contrast to our in vivo observations, a clear induction of a Warburg effect with an increased glycolysis and an inhibited mitochondrial respiration. Together, our study indicates that miR-22-3p is a master regulator of the lipid and glucose metabolism with differential effects in specific organs and in transformed hepatic cancer cells, as compared to non-tumoral tissue.

Abstract 5989: Metabolic heterogeneity of the tumor microenvironment

Abstract Background: Single cell technologies have provided insight into the cellular and phenotypic heterogeneity of the tumor microenvironment (TME). However, major technical hurdles exist to elucidate the metabolic fate of a given metabolite in the cellularly diverse TME. Methods: By collecting tumor interstitial fluid from human kidney tumors and matched normal tissue, our group has gained valuable insight into the metabolic content of the Renal Cell Carcinoma microenvironment and the normal kidney parenchyma. Given the high concentration of glucose in both of these tissue environments (>4 mM), we hypothesized that infiltrating immune cells and transformed cells can use this widely available sugar as a fuel source. To test this hypothesis, we labeled glycolytic cells in the tumor microenvironment using the widely clinically adopted F18 fluoro-2-deoxyglucose (FDG) Positron Emission Tomography (PET) tracer. By conducting successive magnetic bead separation, highly enriched immune cell and non-immune cell fractions were analyzed for radioactive, transcriptomic and metabolic analysis. Results: This work has led to the intriguing discovery that infiltrating CD45+ immune cells are more actively glucose consuming than CD45- tumor cells on a per cell basis. Additionally, even though these cells make up a minority of the population of the entire TME (<15%), they contribute to > 65% of the total glucose uptake in the tumor microenvironment. Subsequent metabolic studies have shown that they are also more actively glycolytic. This phenomenon holds true across multiple orthotopic tumor models in multiple mouse genetic backgrounds. Further characterization of this glycolytic CD45+ fraction illustrates that the CD11b+, F4/80+ tumor associated macrophages (TAM) fraction is the most avid. Additionally, the tumor infiltrating T cells have much higher FDG avidity than the naïve splenocytes and surprisingly the transformed cancer cells. In support of this data, ex vivo seahorse analysis using these fractions demonstrates that the most metabolically active fraction is TAM-rich F4/80+ fraction. Infiltrating CD45+ cells appear to be metabolically fixed, as demonstrated by their lack of glycolytic reserve when compared to transformed tumor cells in the glycostress test. In order to examine the immunogenicity of F18 based tracers, we have traced the fate of F18-Glutamine (Q). Interestingly, the F18-Q accumulates to a much greater extent in the CD45- cells (on a per cell basis), illustrating the anaplerotic need of cancer cells. Conclusion: Ultimately, this work reveals the nutrient environment of the tumor interstitial space at a resolution not previously revealed, and uncovers the diversity of metabolic utilization across cell types in the heterogenous tumor microenvironment. Citation Format: Bradley I. Reinfeld, Matthew Z. Madden, Ahmed Ali, Melissa M. Wolf, Kirsten L. Young, Racheal Hongo, Kathryn E. Beckermann, Matthew G. Vander Heiden, Alex Muir, Jeffrey C. Rathmell, W. Kimryn Rathmell. Metabolic heterogeneity of the tumor microenvironment [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5989.

FCHSD2 controls oncogenic ERK1/2 signaling outcome by regulating endocytic trafficking

The evolution of transformed cancer cells into metastatic tumors is, in part, driven by altered intracellular signaling downstream of receptor tyrosine kinases (RTKs). The surface levels and activity of RTKs are governed mainly through clathrin-mediated endocytosis (CME), endosomal recycling, or degradation. In turn, oncogenic signaling downstream of RTKs can reciprocally regulate endocytic trafficking by creating feedback loops in cells to enhance tumor progression. We previously showed that FCH/F-BAR and Double SH3 Domain-Containing Protein (FCHSD2) has a cancer-cell specific function in regulating CME in non-small-cell lung cancer (NSCLC) cells. Here, we report that FCHSD2 loss impacts recycling of the RTKs, epidermal growth factor receptor (EGFR) and proto-oncogene c-Met (MET), and shunts their trafficking into late endosomes and lysosomal degradation. Notably, FCHSD2 depletion results in the nuclear translocation of active extracellular signal-regulated kinase 1 and 2 (ERK1/2), leading to enhanced transcription and up-regulation of EGFR and MET. The small GTPase, Ras-related protein Rab-7A (Rab7), is essential for the FCHSD2 depletion-induced effects. Correspondingly, FCHSD2 loss correlates to higher tumor grades of NSCLC. Clinically, NSCLC patients expressing high FCHSD2 exhibit elevated survival, whereas patients with high Rab7 expression display decreased survival rates. Our study provides new insight into the molecular nexus for crosstalk between oncogenic signaling and RTK trafficking that controls cancer progression.

Phase I study of SNK01 (autologous non-genetically modified natural killer cells with enhanced cytotoxicity) in refractory metastatic solid tumors.

e15024 Background: Natural Killer (NK) cells possess the innate ability to detect transformed cancer cells and kill them, thus playing a key role in cancer immunosurveillance and antitumor immunity. In general, prior NK cell therapies have not shown efficacy in solid tumors and the expansion of NK cells in cancer patients to clinically therapeutic doses is quite challenging, making allogenic preferable to autologous treatment. SNK01 is a first-in-kind, autologous non-genetically modified NK cell therapy with enhanced cytotoxicity which can be consistently produced even from heavily pretreated cancer patients, and has been shown to have efficacy against numerous solid tumor types in preclinical studies. Methods: In this single-arm Phase I study (NCT03941262) to investigate the safety and efficacy of SNK01, patients with refractory metastatic solid tumors were treated in a 3 + 3 dose escalation study with five weekly infusions of SNK01 at 1, 2, or 4 x 109 cells per infusion. Primary endpoint is safety and quality of life (QoL), and secondary endpoint is objective response rate (ORR). Results: Seven of nine planned patients have been enrolled up to date and five have completed treatment. All patients have rapidly progressive metastatic disease and have received an average of five lines of prior therapy. Tumor types include one non-small lung cancer, one colorectal cancer, and five sarcomas. Median age is 52 (32-62). All patients have had successful expansion and cytotoxic enhancement of their NK cells. Three patients have completed 1 x 109 SNK01 and two patients have completed 2 x 109 SNK01. There have been no adverse events according to NCI-CTCAE v 5.0 or any cytokine release syndrome, and all patients have reported an overall improvement in QoL. At week 9, three of three patients at the 1 x 109 dose and one of two at the 2 x 109 achieved a best overall response of stable disease as per RECIST 1.1. Conclusions: Expanding and increasing the cytotoxicity of NK cells in our heavily pretreated patients has been consistently reliable. SNK01 monotherapy has been very safe and well tolerated in patients with rapidly progressive solid tumors. MTD has not been reached and dose escalation is ongoing. Evaluation of anti-tumor activity is ongoing, but at a minimum, SNK01 appears to slow and possibly halt progression in very aggressive disease and improve quality of life while improving the overall quality of life. The remaining two planned patients are currently being enrolled and a full update will be presented. Clinical trial information: NCT03941262 .

Non-small cell lung cancer-small cell lung cancer transformation as mechanism of resistance to tyrosine kinase inhibitors in lung cancer

Mutated or rearranged driver kinases in non-small cell lung cancer (NSCLC) cells are clinically amenable to treatment with tyrosine kinase inhibitors (TKIs) resulting in prolonged survival and significant benefit compared to cytotoxic chemotherapy. The most frequent genomic alterations are observed for epidermal growth factor receptor and anaplastic lymphoma kinase, which can be blocked by a range of specific TKIs in sequence. In clinics, resistance to TKIs emerges after approximately one year and comprises secondary mutations of the kinases (on-target) or alternative pathways circumventing the original kinase (off-target) alterations. A special feature of NSCLC is the occurrence of histological transformation to small cell lung cancer (SCLC) in up to 14% of cases, which, in general, is accompanied by resistance to the original TKIs. SCLC transformed tumors may be treated with the classical platinum/etoposide regimen but thus far there are no definitive guidelines. Four transformed pleural SCLC lines in our lab indicate the presence of a gradual NSCLC-SCLC shift with overlapping drug sensitivities. In conclusion, the treatment of NSCLC-SCLC transformed cancer cells would need a better chemosensitivity assessment using functional genomics to guide further therapy.

Stopping transformed cancer cell growth by rigidity sensing.

A common feature of cancer cells is the alteration of kinases and biochemical signalling pathways enabling transformed growth on soft matrices, whereas cytoskeletal protein alterations are thought to be a secondary issue. However, we report here that cancer cells from different tissues can be toggled between transformed and rigidity-dependent growth states by the absence or presence of mechanosensory modules, respectively. In various cancer lines from different tissues, cells had over tenfold fewer rigidity-sensing contractions compared with normal cells from the same tissues. Restoring normal levels of cytoskeletal proteins, including tropomyosins, restored rigidity sensing and rigidity-dependent growth. Further depletion of other rigidity sensor proteins, including myosin IIA, restored transformed growth and blocked sensing. In addition, restoration of rigidity sensing to cancer cells inhibited tumour formation and changed expression patterns. Thus, the depletion of rigidity-sensing modules through alterations in cytoskeletal protein levels enables cancer cell growth on soft surfaces, which is an enabling factor for cancer progression.

Human embryonic stem cells display a pronounced sensitivity to the cyclin dependent kinase inhibitor Roscovitine

BackgroundThe essentially unlimited expansion potential and the pluripotency of human embryonic stem cells (hESCs) make them attractive for cell-based therapeutic purposes. Although hESCs can indefinitely proliferate in culture, unlike transformed cancer cells, they are endowed with a cell-intrinsic property termed mitochondrial priming that renders them highly sensitive to apoptotic stimuli. Thus, all attempts to broaden the insights into hESCs apoptosis may be helpful for establishing pro-survival strategies valuable for its in vitro culture and further use in clinical applications. Cyclin-dependent kinases (CDKs), a family of serine/threonine protein kinases originally identified as regulators of the eukaryotic cell cycle, can also regulate transcription and differentiation. Moreover, there are compelling data suggesting that its activities are involved in certain apoptotic programs in different cell types. Currently, it is not completely determined whether CDKs regulate apoptotic processes in rapidly proliferating and apoptosis-prone hESCs. In this study, to elucidate the effect of CDKs inhibition in hESCs we used Roscovitine (ROSC), a purine analogue that selectively inhibits the activities of these kinases.ResultsInhibition of CDKs by ROSC triggers programmed cell death in hESCs but not in proliferating somatic cells (human fibroblasts). The apoptotic process encompasses caspase-9 and -3 activation followed by PARP cleavage. ROSC treatment also leads to p53 stabilization, which coincides with site-specific phosphorylation at serine 46 and decreased levels of Mdm2. Additionally, we observed a transcriptional induction of p53AIP1, a repression of pro-survival factor Mcl-1 and an up-regulation of pro-apoptotic BH3-only proteins NOXA and PUMA. Importantly, we found that the role of CDK2 inhibition appears to be at best accessory as an active CDK2 is not required to ensure hESCs survival.ConclusionOur experimental data reveal that hESCs, contrary to fibroblasts, exhibit a pronounced sensitivity to ROSC.

Abstract 985: Development and validation of cell-based TFEB translocation assay in a high-content and high-throughput screening format

Abstract Transcription factor EB (TFEB) is a master regulator of lysosomal gene expression and mitochondrial biogenesis. Autophagy is known to act as a double edged sword in cancer. Autophagy acts as a tumor suppressor during the tumorigenesis process but it also helps to sustain the survival of already transformed cancer cells. Lysosomes sit at the end of the autophagy pathway, which is critical to meet the needs of autophagy. Therefore, small molecules that inhibit TFEB-mediated lysosomal biogenesis and autophagic degradation may be helpful for inhibiting cancer cell growth and limiting cancer progression. On the other hand, small molecules that enhance TFEB-mediated lysosomal degradation may be used to prevent tumorigenesis. In order to identify compounds that modulate TFEB translocation, an image-based TFEB translocation assay was developed in a 1536-well plate format with a GFP-TFEB stable AML12 cell line, which is an immortalized mouse hepatocyte cell line. Torin 1, a known TFEB translocation inducer, was used as positive control in the study. Torin 1 increased TFEB nuclear translocation in a concentration dependent manner with an EC50 of 150nM. To optimize the assay condition, various cell densities per well and time courses were tested. To validate this assay, a group of 384 known anticancer compounds were screened at 11 concentrations ranging from 0.8 nM to 46 μM. From this screening, we identified 25 compounds that enhanced TFEB nuclear translocation with EC50s ranging from 2 nM to 12 μM. Several known and novel TFEB inducers were among these compounds. These results demonstrate that this high-content and high-throughput assay can be used to screen large numbers of chemicals and provide a robust in vitro system to identify TFEB agonists and antagonists, which may be used to either prevent (agonists) or treat (antagonist) cancer. Citation Format: Li Zhang, Ruili Huang, Wen Xing Ding, Menghang Xia. Development and validation of cell-based TFEB translocation assay in a high-content and high-throughput screening format [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 985.

Effects of PTEN Loss and Activated KRAS Overexpression on Viscoelasticity of Breast Epithelial Cells

Abnormalities in the mechanical properties of cells are associated with disease pathogenesis and progression. In breast cancer, the stiffness of transformed cancer cells are different than healthy breast epithelial cells. The PI3K and Ras/MAPK pathways have been shown to incite cancer dormancy behavior individually and tumorigenic behavior in unison when induced in healthy breast epithelial cells (MCF-10A) in vivo. However, very little is known about the roles of these two pathways in regulating the mechanical properties of cells. We measured the viscoelasticity of MCF-10A cells with PTEN (phosphatase and tensin homolog) knockout and activated KRAS (Kristen rat sarcoma viral oncogene homolog) overexpression to activate the PI3K and Ras/MAPK pathways with atomic force microscopy. Our results suggest that PTEN knockout alone resulted in a decrease in cell stiffness and increase in fluid-like behavior. Activated KRAS overexpression increased cell stiffness regardless of PTEN expression level.

Normalizing Transformed Cancer Cells with Rigidity Sensing

Normalizing Transformed Cancer Cells with Rigidity Sensing

Remodeling the Microenvironment before Occurrence and Metastasis of Cancer.

Tumorigenesis and progression of cancer are complex processes which transformed cells and stromal cells interact and co-evolve. Intrinsic and extrinsic factors cause the mutations of cells. The survival of transformed cells critically depends on the circumstances which they reside. The malignant transformed cancer cells reprogram the microenvironment locally and systemically. The formation of premetastatic niche in the secondary organs facilitates cancer cells survival in the distant organs. This review outlines the current understanding of the key roles of premalignant niche and premetastatic niche in cancer progression. We proposed that a niche facilitates survival of transformed cells is characteristics of senescence, stromal fibrosis and obese microenvironment. We also proposed the formation of premetastatic niche in secondary organs is critically influenced by primary cancer cells. Therefore, it suggested that strategies to target the niche can be promising approach to eradicate cancer cells.

Actin Dysfunction Induces Cell Cycle Delay at G2/M with Sustained ERK and RSK Activation in IMR-90 Normal Human Fibroblasts.

The actin cytoskeleton plays a key role in the entry of mitosis as well as in cytokinesis. In a previous study, we showed that actin disruption delays mitotic entry at G2/M by sustained activation of extracellular signal-related kinase 1/2 (ERK1/2) in primary cells but not in transformed cancer cell lines. Here, we examined the mechanism of cell cycle delay at G2/M by actin dysfunction in IMR-90 normal human fibroblasts. We observed that de-polymerization of actin with cytochalasin D (CD) constitutively activated ribosomal S6 kinase (RSK) and induced inhibitory phosphorylation of Cdc2 (Tyr 15) in IMR-90 cells. In the presence of an actin defect in IMR-90 cells, activating phosphorylation of Wee1 kinase (Ser 642) and inhibitory phosphorylation of Cdc25C (Ser 216) was also maintained. However, when kinase-dead RSK (DN-RSK) was over-expressed, we observed sustained activation of ERK1/2, but no delay in the G2/M transition, demonstrating that RSK functions downstream of ERK in cell cycle delay by actin dysfunction. In DN-RSK overexpressing IMR-90 cells treated with CD, phosphorylation of Cdc25C (Ser 216) was blocked and phosphorylation of Cdc2 (Tyr 15) was decreased, but the phosphorylation of Wee1 (Ser 642) was maintained, demonstrating that RSK directly controls phosphorylation of Cdc25C (Ser 216), but not the activity of Wee1. These results strongly suggest that actin dysfunction in primary cells activates ERK1/2 to inhibit Cdc2, delaying the cell cycle at G2/M by activating downstream RSK, which phosphorylates and blocks Cdc25C, and by directly activating Wee1.

Transcriptome profiling of the interconnection of pathways involved in malignant transformation and response to hypoxia.

In tumor tissues, hypoxia is a commonly observed feature resulting from rapidly proliferating cancer cells outgrowing their surrounding vasculature network. Transformed cancer cells are known to exhibit phenotypic alterations, enabling continuous proliferation despite a limited oxygen supply. The four-step isogenic BJ cell model enables studies of defined steps of tumorigenesis: the normal, immortalized, transformed, and metastasizing stages. By transcriptome profiling under atmospheric and moderate hypoxic (3% O2) conditions, we observed that despite being highly similar, the four cell lines of the BJ model responded strikingly different to hypoxia. Besides corroborating many of the known responses to hypoxia, we demonstrate that the transcriptome adaptation to moderate hypoxia resembles the process of malignant transformation. The transformed cells displayed a distinct capability of metabolic switching, reflected in reversed gene expression patterns for several genes involved in oxidative phosphorylation and glycolytic pathways. By profiling the stage-specific responses to hypoxia, we identified ASS1 as a potential prognostic marker in hypoxic tumors. This study demonstrates the usefulness of the BJ cell model for highlighting the interconnection of pathways involved in malignant transformation and hypoxic response.

MP81-15 PROSTATE SMALL CELL CARCINOMA EXPRESS STEM CELL FACTORS THAT CAN TRANSFORM NORMAL AND CANCER CELLS INTO STEM-LIKE

You have accessJournal of UrologyStem Cell Research: Stem Cell Research II1 Apr 2018MP81-15 PROSTATE SMALL CELL CARCINOMA EXPRESS STEM CELL FACTORS THAT CAN TRANSFORM NORMAL AND CANCER CELLS INTO STEM-LIKE Adelle Kanan and Alvin Liu Adelle KananAdelle Kanan More articles by this author and Alvin LiuAlvin Liu More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.2723AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Prostate cancer can be grouped into either luminal-like (differentiated) or stem-like (undifferentiated). The latter type includes small cell carcinoma. The LuCaP 145.1 small cell carcinoma xenograft line shows expression of stem cell transcription factors LIN28A, NANOG, POU5F1 and SOX2. Luminal-like adenocarcinoma LuCaP lines show expression of only one or two of these factors, while cancer cells of Gleason patterns 3 and 4 do not. Forced expression of these four genes in differentiated somatic cells produces induced pluripotent stem (iPS) cells. We want to determine if other LuCaP small cell carcinoma xenografts also express these genes, and whether these genes cloned from LuCaP 145.1 can also transform cell types, both normal and cancer, into stem-like. METHODS LuCaP small cell carcinoma harvested from mice were minced and digested by collagenase. The resultant cells were centrifuged on Percoll density gradients. Gene expression was analyzed by RT-PCR. Full-length LIN28A, NANOG, POU5F1 and SOX2 cDNA (Kozak box, ATG start to stop codon) were cloned from LuCaP 145.1 into mammalian expression vector pVITRO1-neo, and transfected by electroporation into HEK293F fibroblasts and luminal-like prostate cancer cells LNCaP and C4-2B. Individual colonies under drug G418 selection (1 mg/ml) were cloned, and analyzed for gene expression. Transfected cells were grown 1) in media supplemented with 10% fetal bovine serum under normoxia, 2) in serum-free media with KnockOut Serum Replacement, 3) on Matrigel-coated plates, 4) on irradiated mouse embryonic fibroblasts (MEF) under hypoxia. RESULTS The four stem cell factor genes were expressed by all established LuCaP small cell carcinoma examined: LuCaP 93, 145.1, 145.2, 173.1, 173.2A. LuCaP 145.1 cells also showed a stromal cell density (1.035). Transfection of plasmid pLP2 containing LIN28A/POU5F1 and pSN4 containing SOX2/NANOG into HEK293F, LNCaP and C4-2 produced cells displaying a stem cell-like colony morphology. Neo (G418)-resistant cells appeared in ~ 1 week, and showed expression of the stem cell factor genes as well as low expression of B2M (β 2-microglobulin) a characteristic of stem cells. The colony morphology was distinct from that of the untransfected cells, and showed differences among the culture conditions used. CONCLUSIONS Transcription factors cloned from small cell carcinoma are capable of transforming normal and cancer cells. The generated plasmid vectors are safer to use than lentiviral vectors; G418 allows selection of transfected cells, and deters microbial contamination. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e1102 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Adelle Kanan More articles by this author Alvin Liu More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Diethylamino-curcumin mimic with trizolyl benzene enhances TRAIL-mediated cell death on human glioblastoma cells

Backgrounds Glioblastoma multiforme is one of the most aggressive human malignant brain tumors. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is known as the death ligand, which induces preferential apoptosis of transformed cancer cells. In this study, we demonstrated that the newly synthesized diethylamino-curcumin mimic with trizolyl benzene (YM-4) enhances cytotoxicity in combination with TRAIL in human glioblastoma cells.

Algorithms of the Femtoscope: KeV X-Rays Cure Cancer While MeV X-Rays Only Burn the Cells

The study of cancer with the Femtoscope shows us that the information of the cell nucleus is correlated with the atomic nucleus. Femtoscope and entropy algorithms monitor the time and energy of x-rays that transform cancer cells into healthy cells and vice versa. Curing cancer means recovering the information lost from a cancer cell, leading to a minimum entropy. The efficient treatment of cancer presents resonance frequencies in the production and elimination of cancerous cells, asymmetrically. The cure asymmetry of cancer is due to the support of DNA repair genes, allowing the stability of a race or species, and prioritizing life to death. Using the Femtoscope and Spectroscopy, we experimentally validate the resonance frequencies, which effectively cure the cancer and find the optimal times and doses of treatment. In this way, we minimize collateral effects and unnecessary economic costs. In addition, the phosphorus resonance demonstrates why the low energies of x-rays cure cancer and high x-ray energies only burn cancer cells.

The Carcinogenic Potential of Microbial Infections of the Prostate and the Role of Contaminations

Several studies have identified bacteria and viruses in human prostatic tissues. The tumor microenvironment of prostate carcinoma is a complex community of genetically transformed cancer cells, non-neoplastic cells, and a diverse collection of microorganisms. Each of these components may contribute to carcinogenesis; however, the role of the microbes is the least well known. A variety of detection techniques have been used, such as PCR-based approaches, fluorescence in situ hybridization, immunological detection assays, and (bacterial) cultivation. Detection rates vary between these methods and each method has specific advantages and limitations. However, confounded by the high risk of contamination during or after the biopsy, it is challenging to make solid conclusions about the presence of certain microorganisms and its possible role in disease formation or progression. This doubt increases with the sensitive detection methods such as next generation sequencing technologies if there are no proper controls. Resident microbial communities often differ between healthy and diseased states, but whether these differences are of primary etiological importance or secondary to the altered inflammatory environment remains largely unknown.

A vimentin binding small molecule leads to mitotic disruption in mesenchymal cancers

Expression of the transcription factor FOXC2 is induced and necessary for successful epithelial-mesenchymal transition, a developmental program that when activated in cancer endows cells with metastatic potential and the properties of stem cells. As such, identifying agents that inhibit the growth of FOXC2-transformed cells represents an attractive approach to inhibit chemotherapy resistance and metastatic dissemination. From a high throughput synthetic lethal screen, we identified a small molecule, FiVe1, which selectively and irreversibly inhibits the growth of mesenchymally transformed breast cancer cells and soft tissue sarcomas of diverse histological subtypes. FiVe1 targets the intermediate filament and mesenchymal marker vimentin (VIM) in a mode which promotes VIM disorganization and phosphorylation during metaphase, ultimately leading to mitotic catastrophe, multinucleation, and the loss of stemness. These findings illustrate a previously undescribed mechanism for interrupting faithful mitotic progression and may ultimately inform the design of therapies for a broad range of mesenchymal cancers.

The Regulatory Role of IFN-γ on the Proliferation and Differentiation of Hematopoietic Stem and Progenitor Cells.

The replenishment of all blood cell lineages is hierarchically organized by the process of hematopoiesis, which is based on the differentiation pathways of hematopoietic stem and progenitor cells (HSPCs). Due to the ability to balance between self-renewal and differentiation, hematopoietic stem cells (HSCs) can generate the appropriate cell type that is required by the immune system and peripheral blood in response to physiological or pathological conditions. Numerous studies have shown that some proinflammatory cytokines contribute to the regulation of the various hematopoietic compartments. Of these, IFN-γ is a type II interferon primarily produced by T cells and natural killer cells, and plays a major role in the defense against invading pathogens and transformed cancer cells; moreover, a growing amount of research indicates that it exerts negative or positive regulatory effect on hematopoiesis. Although IFN-γ is a widely regarded negative regulator of HSC proliferation, it also participates in some chronic infections or hematological malignancies that induce bone marrow failure. Recent studies have demonstrated unexpected effects of IFN-γ, including the promotion of HSC formation and the stimulation of myelopoiesis. Here, we review the direct and indirect effects of IFN-γ on hematopoiesis, as well as the underlying signaling mechanisms of how IFN-γ modulates the self-renewal, cell cycle entry, and proliferation of HSCs. Next, we describe how IFN-γ affects different stages of the lineage differentiation from HSCs. Finally, we discuss the relationship between IFN-γ and compensatory extramedullary hematopoiesis, as well as some related clinical diseases.

Abstract 5458: Oncostatin M induces epithelial-mesenchymal transition and stem cell properties in senescent human mammary epithelial cells

Abstract The Interleukin-6 (IL-6) cytokine family member Oncostatin M (OSM) induces epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC) properties in transformed cancer cells. Paradoxically, OSM induces a growth-inhibiting senescence response in normal human mammary epithelial cells (HMEC). Senescence is induced in normal cells by various stresses, such as aberrant oncogene signaling, and serves as a major tumor-suppressive barrier that must be bypassed for cellular transformation in vitro and tumorigenesis in vivo. Previous reports show that aberrant reactivation of EMT-inducing transcription factors (EMT-TFs) promotes senescence bypass. Thus, aberrant EMT may drive tumor initiation in addition to providing cancer cells with mesenchymal cell and stem cell properties that drive tumor progression, metastasis, and tumor recurrence. Conversely, our studies here indicate that much like aberrant oncogene signaling, aberrant EMT and CSC-properties induced by OSM may actually engage senescence in normal cells. Infecting Primary HMEC isolated from reduction mammoplasty tissue with lentivirus encoding shRNA to p16 created indefinite lifespan “Normal” HMEC. Subsequent infections with viruses encoding shRNA to p53, then constitutive expression of MYC and RAS created “Transformed” HMEC. After treating HMEC with recombinant OSM, proliferation and senescence were assessed by growth assays and qRT-PCR analysis of senescence-associated β-galactosidase (GLB1) expression. EMT and CSC-properties were assessed by western and qRT-PCR analysis of an epithelial marker E-cadherin (CDH1), a mesenchymal marker Vimentin (VIM), the EMT-TF Snail (SNAI1), and the CSC markers CD44 and HAS2. Flow cytometry examining BrdU-incorporation and CD24/CD44 surface expression also assessed proliferation and a CSC phenotype. OSM reduced growth by 80% and induced GLB1 expression in Normal HMEC, but not in Transformed HMEC. OSM-induced senescence was accompanied by Snail induction, changes in EMT-associated markers (reduced E-cadherin/increased Vimentin), and CSC marker (CD44 and HAS2) induction. Notably, OSM induced immediate SNAI1 and HAS2 expression; and a majority of the OSM-induced CD44+ population consisted of non-proliferating (BrdU-) cells. Unexpectedly, Snail and CD44 were also induced during oncogenic RAS-induced senescence; and Snail expression alone reduced growth and induced CD44 in normal HMEC. These results suggest that much like aberrant oncogene signaling, aberrantly induced EMT and CSC-properties may engage senescence in normal cells rather than promoting senescence bypass. Defining the mechanism linking senescence, EMT, and CSC-properties will identify targets for novel “pro-senescence” therapies that may inhibit metastasis and tumor recurrence, and expand our repertoire of therapies that enhance cancer patient outcomes. Citation Format: Benjamin L. Bryson, Damian J. Junk, Mark W. Jackson. Oncostatin M induces epithelial-mesenchymal transition and stem cell properties in senescent human mammary epithelial cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5458. doi:10.1158/1538-7445.AM2017-5458