Publications

The Roles of Ik(Beta) Kinases in Prostate Carcinogenesis and the Effect of Their Inhibition on Survival of Prostate Tumors

Abstract : This research project investigates the role of the IkB kinase (IKK) and NF-kB in the development of prostate cancer (CaP), and examines the possibility that IKK inhibitors can be used in CaP treatment. To reach this goal we are employing two mouse models in which either the IKK(beta) or the IKK(alpha) subunit of IKK are deleted or inhibited in prostate epithelial cells. We found that neither IKK(beta) nor IKK(alpha) are required for normal prostate development, however IKK(alpha) may play an important role in the development of advanced CaP. The study on the role of IKK(beta) in prostate carcinogenesis in animal models is ongoing. We also found that IKK/NF-kB activities were increased during the evolution of androgen-independent CaP, a response that could be mediated by some of androgen-regulated genes, such as TMFF2. Furthermore, we found that a prototypical IKK inhibitor IT-3 can suppress the proliferation of human CaP cells. Although a more thorough examination of the role of IKK/NF-kB in CaP development and progression is currently underway, our results obtained during last year suggest an important role for both IKK(alpha and IKK(beta) in development and progression of CaP. Therefore inhibition of either protein kinase or both would be an effective and attractive option for the treatment of CaP.

The Fire within: Cell-Autonomous Mechanisms in Inflammation-Driven Cancer

Compliance with physical activity guidelines : prevalence in a population of children and youth

No abstract is provided for this article.

Abstract 3: Imaging of stromal cells during hepatocellular metastasis formation in the RFP-MUP-uPA mouse

Abstract Major urinary protein-urokinase type plasminogen activator (MUP-uPA) transgenic mice develop liver disease. We have developed the RFP-MUP-uPA mice, by crossing MUP-uPA mice with transgenic mice ubiquitously-expressing RFP. Hepatocyte RFP fluorescence in this model is very strong. In addition, RFP fluorescence of stromal cells is very bright enabling the portal vein and sinusoid structures to be clearly visualized. Huh7 liver cancer cells expressing green fluorescent protein (GFP) were injected in the spleen of MUP-uPA transgenic mice. The vessels of the MUP-uPA liver are filled with Huh7-GFP cells as imaged in live mice 3h after cell injection. Huh7-GFP cells formed tumor colonies in the liver 28 days after cell transplantation to the spleen. The Huh7-GFP cells were visualized in the liver with the Olympus FV1000 confocal microscope. In large metastatic liver tumors in RFP-MUP-uPA mice, stroma cells derived from the host animal appeared around and into the edge of Huh7-GFP tumors. In contrast, in small metastatic liver tumors, there were no stromal cells observed. The stromal cells were observed in the spleen primary tumor, as well as kidney and the lung metastasis in addition to the large liver tumors. This model enables imaging the effects of uPA on liver metastasis and stromal recruitment enabling the metastasis to progress. Citation Format: Atsushi Suetsugu, Hisanobu Ogata, Yukihiko Hiroshima, Masashi Momiyama, Yosuke Osawa, Hisataka Moriwaki, Shigetoyo Saji, Michael Karin, Robert M. Hoffman. Imaging of stromal cells during hepatocellular metastasis formation in the RFP-MUP-uPA mouse. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3. doi:10.1158/1538-7445.AM2014-3

Abstract 3528: Influence of obesity-associated intra-tumor microbes on exacerbating cancer severity

Abstract Background: Despite there being a well-established connection between the gut microbiome and metabolic disease in humans, intra-tumor microbes and the mechanisms by which they regulate cell signaling, inflammation, and adipocyte growth to exacerbate disease severity in cancer patients also suffering from obesity remains largely unclear. In this study, we identified microbes to be distinctly abundant in cancer patients who are obese and correlated microbe abundance to patient survival, clinical variables, and immunological genes and pathways, in order to mechanistically explain how differential microbe abundance may influence clinical outcome. Methods: Microbial reads were aligned and extracted from raw whole-transcriptome RNA-sequencing data of cancer patient samples using Pathoscope 2.0 software. The Kruskal-Wallis test was used to correlate body mass index (BMI)-associated microbes to clinical variables. Reactome FIViz and Gene Set Enrichment Analysis were used to calculate pathway enrichment. Results: We identified specific microbes, including Pseudomonas fluorescens SBW25 and Enterobacter cloacae, and chemokine and interleukin-related genes to be potential determining factors of disease severity among cancer patients in BMI-associated groups. Gene set enrichment analysis revealed that microbes abundant in cancer tissue in obese patient samples, including Pseudomonas baetica in liver cancer patients, were significantly associated with the upregulation oncogenic, cell migration-related signaling pathways. Intra-tumor microbes from obese patient samples were also found to correlate with chemokine signaling and TFR2/NFkB-related genes, both of which have well-established roles in inflammatory activity. Conclusions: Our study significantly advances the understanding of the microbiome composition of the tumor microenvironment in patients who are obese and microbes’ relationship with clinical and immunologic variables, particularly inflammatory-related genes and pathways. We uncovered unknown mechanisms of the microbiome-immune interaction and obtained definitive data on microbiome dysbiosis in patients with obesity as a key determinant of severity of cancer, including microbe regulation of inflammasome activity. While deeper sequencing, more rigorous contamination correction, and in vitro and in vivo experiments are necessary to fully elucidate how microbe species can effectively act as therapeutic agents in probiotic and prebiotic therapies to reduce insulin resistance, inflammation, and glucose levels, our results are essential for guiding this future research. Citation Format: Aditi Gnanasekar, Neil Shende, Jaideep Chakladar, Wei T. Li, Lindsay M. Wong, Michael Karin, Weg M. Ongkeko. Influence of obesity-associated intra-tumor microbes on exacerbating cancer severity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3528.

Abstract 976: Junctional adhesion molecule-A (JAM-A) expression is downmodulated by miR-21 during colorectal cancer progression

Abstract Introduction: MicroRNAs are small non-coding-RNAs that control cell homeostasis and are deregulated in human colorectal cancer (CRC). MiR-21 up-regulation is frequent in CRC and represents a driver for tumor initiation and progression. Junctional Adhesion Molecule A (JAM-A) is a tight junction protein involved in regulation of para-cellular permeability in epithelial and endothelial cells. JAM-A forms homodimers and activates signalling pathways regulating cell adhesion, polarity and proliferation. For this reason, monoclonal antibodies against JAM-A are currently tested in the preclinical setting as potential targeted therapies. JAM-A loss appears to be an early event in CRC initiation but very little is known about the role of JAM-A in human sporadic CRC. The aim of our study was defining the pattern of expression of JAM-A in human CRC and to the define the whether its deregulation is a consequence of aberrant microRNA expression. Methods: JAM-A expression was analyzed by immunohistochemistry in 1254 cases of human sporadic CRC. MiR-21 and JAM-A expression was analyzed in human CRC cells [RKO, HT29, HCT116, SW620, HCT15, SW480, SW837, SW48, DLD1 (both Wild-Type and Knock-Out for miR-21)] by Real-Time-PCR and Western-Blotting. MiR-21 silencing and over-expression were performed using Exiqon Locked Nucleic Acids and Ambion Pre-miR respectively. MiR-21 binding site in JAM-A 3’UTR was predicted using bio-informatics tools (RNA Hybrid) and a 236bp area encompassing the predicted seed region was cloned downstream of the Firefly Luciferase gene in PGL3 vector for Luciferase Reporter Assay. Results: JAM-A down-regulation was observed in cancer compared to normal adjacent tissues. JAM-A was progressively down-regulated in the progression from normal epithelium, dysplasia, intraepithelial and invasive cancer. Poorly differentiated cancers showed total loss of JAM-A. Shifted staining from apical to baso-lateral compartment was observed in 25% of all cancers. Silencing and re-expression of miR-21 in CRC cell lines resulted in increase and down-regulation of JAM-A protein expression respectively. Luciferase reporter assay experiments were able to define the potential seed region by which miR-21 interacts with JAM-A 3′UTR. Conclusion: Our data suggest that JAM-A expression is deregulated during human colorectal cancer progression as a consequence of miR-21 over-expression. These results highlight how miR-21 could potentially exert its’ actions on para-cellular permeability and cell polarity acting on junctional adhesion molecules. Understanding the mechanisms of JAM-A loss or re-localization may help to stratify patient's treatment and identify patients who might benefit from anti-JAM-A monoclonal antibodies. Citation Format: Andrea Lampis, Claudio Murgia, Viola Paulus-Hock, Matteo Fassan, Joanne Edwards, Paul Horgan, Owen Sansom, Luigi Terracciano, Michael Karin, Carlo Croce, Chiara Braconi, Nicola Valeri. Junctional adhesion molecule-A (JAM-A) expression is downmodulated by miR-21 during colorectal cancer progression. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 976. doi:10.1158/1538-7445.AM2014-976

Psychosocial correlates of dietary intake among overweight and obese men.

To investigate the relationship between theoretically based psychosocial constructs and dietary components among overweight men.Participants were 441 men (BMI M = 34.2). Psychosocial constructs included self-efficacy, decisional balance, social support, and behavior change strategies. Dietary components were fat, fiber, and fruit and vegetable intake.All significant findings were in the expected direction. Multiple regression models indicated that the psychosocial factors accounted for the most variance in vegetable intake (R(2)=.13) and the least variance in fat (R(2)=.05).Theoretically based psychosocial constructs were related to overweight men's dietary intake and have potential for use in tailored behavior-change interventions.

Environmental Correlates of Physical Activity

A Novel Tetanus Neurotoxin-insensitive Vesicle-associated Membrane Protein in SNARE Complexes of the Apical Plasma Membrane of Epithelial Cells

The importance of soluble N-ethyl maleimide (NEM)-sensitive fusion protein (NSF) attachment protein (SNAP) receptors (SNAREs) in synaptic vesicle exocytosis is well established because it has been demonstrated that clostridial neurotoxins (NTs) proteolyze the vesicle SNAREs (v-SNAREs) vesicle-associated membrane protein (VAMP)/brevins and their partners, the target SNAREs (t-SNAREs) syntaxin 1 and SNAP25. Yet, several exocytotic events, including apical exocytosis in epithelial cells, are insensitive to numerous clostridial NTs, suggesting the presence of SNARE-independent mechanisms of exocytosis. In this study we found that syntaxin 3, SNAP23, and a newly identified VAMP/brevin, tetanus neurotoxin (TeNT)-insensitive VAMP (TI-VAMP), are insensitive to clostridial NTs. In epithelial cells, TI-VAMP-containing vesicles were concentrated in the apical domain, and the protein was detected at the apical plasma membrane by immunogold labeling on ultrathin cryosections. Syntaxin 3 and SNAP23 were codistributed at the apical plasma membrane where they formed NEM-dependent SNARE complexes with TI-VAMP and cellubrevin. We suggest that TI-VAMP, SNAP23, and syntaxin 3 can participate in exocytotic processes at the apical plasma membrane of epithelial cells and, more generally, domain-specific exocytosis in clostridial NT-resistant pathways.

Lack of Physical Activity Associated with Higher Rates of Severe COVID-19 Outcomes

Background: Of the studies examining risk factors for severe COVID-19 outcomes, none have examined inadequate physical activity (PA). We compared hospitalization rates, intensive care unit (ICU) admissions and mortality for COVID-19 patients who were inactive, inconsistently active, or regularly physically active.Methods: This retrospective observational study was done at Kaiser Permanente in Southern California. We identified 48,440 adult patients with a COVID-19 diagnosis from January 1, 2020 to October 21, 2020, with at least two Exercise Vital Sign (EVS) measurements from March 19, 2018 to March 18, 2020. We linked each patient's self-reported PA category (consistently inactive = 0-10 min/week, inconsistently active = 11-149 min/week, consistently active = 150+ min/week) to risk of hospitalization, ICU admission and death after COVID-19 diagnosis. We conducted multivariate logistic regression controlling for known risk factors to assess whether regular PA was associated with COIVD -19 outcomes.Findings: COVID-19 patients who were consistently inactive were more likely to be hospitalized (OR 2.26; 95% CI: 1.81, 2.83), admitted to the ICU (OR= 1.73; 95% CI: 1.18, 2.55) and die (OR 2.49; 95% CI: 1.33, 4.67) than patients who were consistently active. Patients who were inconsistently active were more likely to be hospitalized (OR 1.89; 95% CI: 1.53, 2.33), admitted to the ICU (OR 1.58; 95% CI: 1.10, 2.27) and die (OR 1.88; 95% CI: 1.02, 3.47) compared to those who were consistently active. Patients who were consistently inactive had a greater risk of being hospitalized (OR= 1.20; 95% CI: 1.10, 1.32), admitted to the ICU (OR 1.10; 95% CI: 0.93, 1.29) and death (OR 1.32; 95% CI: 1.09, 1.60) than patients who were inconsistently active.Interpretations: Regular PA is a strong, modifiable independent risk factor for protecting against severe COVID-19 outcomes. Pandemic control measures should prioritize supporting regular PA to lessen COVID-19 morbidity and mortality.Funding Statement: This study was in part supported through funding from the Kaiser Permanente Community Benefits Funds.Declaration of Interests: The authors report no conflict of interest except Dr. Tartof reports grants from Roche/Genentech, Inc., during the conduct of the study.Ethics Approval Statement: The study was approved by the KPSC Institutional Review Board.

EFFECTS OF A PHYSICAL ACTIVITY PROMOTION COURSE FOR UNIVERSITY SENIORS: PROJECT GRAD 897

Project GRAD (Graduate Ready for Activity Daily) is a randomized controlled study of a course for university seniors designed to promote adoption and maintenance of physical activity in preparation for the transition after graduation. A total of 185 women and 153 men were recruited, randomized to intervention or control conditions, and completed baseline and 3-month post-test measures. The intervention consisted of a 2-unit course with lecture and lab components. Lectures covered exercise science and behavior change principles. Weekly labs were led by trained peers and featured supervised physical activity and application of behavior change techniques. The 2-unit control course consisted of lectures on a variety of health topics. Physical activity was assessed at pre- and post-test with telephone administered 7-day physical activity recall interviews that were modified to include reports of exercises for strength and flexibility. Data were analyzed separately for women and men using a 2×2×2 repeated measures ANCOVA (condition× active/inactive at baseline × time), with season as a covariate. There was a significant intervention effect (increase) on total leisure physical activity for women (p<.02), but no effect for men. Although results were in favorable directions, there were no significant intervention effects on leisure-time moderate or vigorous physical activity for either sex. On reported minutes of both strengthening (p<.001) and flexibility(p<.01) exercises, there were significant intervention effects (increases) for women only. The health-related physical activity course was effective for women but not for men. The course increased physical activity similarly for women who were initially regularly active and those who were relatively inactive. Intervention is continuing to promote maintenance of physical activity after graduation.

SupplementaryFigure from Concurrent Disruption of the Ras/MAPK and NF-κB Pathways Induces Circadian Deregulation and Hepatocarcinogenesis

SupplementaryFigure from Concurrent Disruption of the Ras/MAPK and NF-κB Pathways Induces Circadian Deregulation and Hepatocarcinogenesis

The Chemokine CXCL16 and Its Receptor, CXCR6, as Markers and Promoters of Inflammation-Associated Cancers

Clinical observations and mouse models have suggested that inflammation can be pro-tumorigenic. Since chemokines are critical in leukocyte trafficking, we hypothesized that chemokines play essential roles in inflammation-associated cancers. Screening for 37 chemokines in prostate cancer cell lines and xenografts revealed CXCL16, the ligand for the receptor CXCR6, as the most consistently expressed chemokine. Immunohistochemistry and/or immunofluorescence and confocal imaging of 121 human prostate specimens showed that CXCL16 and CXCR6 were co-expressed, both on prostate cancer cells and adjacent T cells. Expression levels of CXCL16 and CXCR6 on cancer cells correlated with poor prognostic features including high-stage and high-grade, and expression also correlated with post-inflammatory changes in the cancer stroma as revealed by loss of alpha-smooth muscle actin. Moreover, CXCL16 enhanced the growth of CXCR6-expressing cancer and primary CD4 T cells. We studied expression of CXCL16 in an additional 461 specimens covering 12 tumor types, and found that CXCL16 was expressed in multiple human cancers associated with inflammation. Our study is the first to describe the expression of CXCL16/CXCR6 on both cancer cells and adjacent T cells in humans, and to demonstrate correlations between CXCL16 and CXCR6 vs. poor both prognostic features and reactive changes in cancer stoma. Taken together, our data suggest that CXCL16 and CXCR6 may mark cancers arising in an inflammatory milieu and mediate pro-tumorigenic effects of inflammation through direct effects on cancer cell growth and by inducing the migration and proliferation of tumor-associated leukocytes.

Cross-validation of the factorial structure of the Neighborhood Environment Walkability Scale (NEWS) and its abbreviated form (NEWS-A)

The Neighborhood Environment Walkability Scale (NEWS) and its abbreviated form (NEWS-A) assess perceived environmental attributes believed to influence physical activity. A multilevel confirmatory factor analysis (MCFA) conducted on a sample from Seattle, WA showed that, at the respondent level, the factor-analyzable items of the NEWS and NEWS-A measured 11 and 10 constructs of perceived neighborhood environment, respectively. At the census blockgroup (used by the US Census Bureau as a subunit of census tracts) level, the MCFA yielded five factors for both NEWS and NEWS-A. The aim of this study was to cross-validate the individual- and blockgroup-level measurement models of the NEWS and NEWS-A in a geographical location and population different from those used in the original validation study. A sample of 912 adults was recruited from 16 selected neighborhoods (116 census blockgroups) in the Baltimore, MD region. Neighborhoods were stratified according to their socio-economic status and transport-related walkability level measured using Geographic Information Systems. Participants self-completed the NEWS. MCFA was used to cross-validate the individual- and blockgroup-level measurement models of the NEWS and NEWS-A. The data provided sufficient support for the factorial validity of the original individual-level measurement models, which consisted of 11 (NEWS) and 10 (NEWS-A) correlated factors. The original blockgroup-level measurement model of the NEWS and NEWS-A showed poor fit to the data and required substantial modifications. These included the combining of aspects of building aesthetics with safety from crime into one factor; the separation of natural aesthetics and building aesthetics into two factors; and for the NEWS-A, the separation of presence of sidewalks/walking routes from other infrastructure for walking. This study provided support for the generalizability of the individual-level measurement models of the NEWS and NEWS-A to different urban geographical locations in the USA. It is recommended that the NEWS and NEWS-A be scored according to their individual-level measurement models, which are relatively stable and correspond to constructs commonly used in the urban planning and transportation fields. However, prior to using these instruments in international and multi-cultural studies, further validation work across diverse non-English speaking countries and populations is needed.

New insights into the pathogenesis and treatment of non-viral hepatocellular carcinoma: a balancing act between immunosuppression and immunosurveillance

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. HCC initiates as a consequence of chronic liver damage and inflammation caused by hepatitis B and C virus infections, excessive alcohol consumption, or non-alcoholic fatty liver disease (NAFLD). Until recently, no effective treatments for advanced HCC were available and the 5-year survival rate had remained below 8% for many years. New insights into the mechanisms that drive the development of NAFLD-related HCC indicate that loss of T-cell-mediated immunosurveillance plays a cardinal role in tumor growth and malignant progression, in addition to previously identified inflammation-driven compensatory proliferation. Recently completed groundbreaking clinical studies have shown that treatments that restore antitumor immunity represent a highly effective therapeutic option for approximately 20% of advanced HCC patients. Understanding the causes of inflammation-driven immunosuppression and immune system dysfunction in the 80% of patients who fail to reignite antitumor immunity despite treatment with checkpoint inhibitors should lead to further and even more dramatic improvements in HCC immunotherapy.

Microscale Audit of Pedestrian Streetscapes--Global

Innate Immunity Gone Awry: Linking Microbial Infections to Chronic Inflammation and Cancer

Habitual Physical Activity of Classroom Teachers: Does It Relate to Their Conduct of Physical Education Classes?.

This study used systematic, direct observation of classes over 2 years to investigate whether physical education (PE) teachers' habitual physical activity related to their conduct of PE classes. Participants were 18 fourth and fifth grade classroom teachers employed in a California school district where classroom teachers typically were responsible for teaching their own PE. They had participated in staff development and received a PE curriculum, and they were encouraged to be physically active for their own health. Over the 4 semesters, teachers completed a self-report instrument that examined how many times they did bouts of strenuous, moderate, and mild physical activity/exercise for 15 minutes or more during their free time the previous week. During the same 4 semesters, researchers observed the teachers' PE classes systematically using the System for Observing Fitness Instruction Time instrument. Data analysis indicated that teachers with greater habitual physical activity promoted fitness more than their less active counterparts. They were much more actively engaged in providing instruction, which indicates a higher quality of instruction. The more active teachers tended to provide more opportunities for their students to participate in moderately vigorous physical activity and vigorous activity bouts. (Contains 15 references.) (SM) ******************************************************************************** Reproductions supplied by EDRS are the best that can be made from the original document. ******************************************************************************** Habitual physical activity of classroom teachers: Does it relate to their conduct of physical education classes? Kathryn LaMaster Thom McKenzie Simon Marshall James Sallis San Diego State University