KRAS est parmi les gènes les plus fréquemment mutés dans les cancers humains, tel que ~ 45% des cancers colorectaux (CCR). Malgré les efforts déployés pour réduire son potentiel oncogénique, KRAS muté est fréquemment associé à la résistance aux médicaments et est extrêmement difficile à cibler sur le plan thérapeutique. Les protéines à la surface cellulaire sont souvent dérégulées dans les cancers et sont des cibles thérapeutiques attrayantes en raison de leur accessibilité aux anticorps. Nous avons séquençé les ARNm de cellules épithéliales intestinales exprimant KRAS muté et observé que ces dernières présentaient des changements importants dans les gènes codant pour des protéines de surface cellulaire. Par conséquent, notre objectif était d'identifier de nouvelles cibles thérapeutiques exprimées à la surface de cellules transformées par l’oncogène KRAS. En utilisant une approche de pointe en protéomique de surface cellulaire, nous avons identifié plusieurs protéines différentiellement exprimées dans les cellules avec KRAS muté par rapport à leurs homologues de type sauvage. Nous avons ensuite effectué un crible CRISPR/Cas9 basé sur les protéines de surface cellulaire, qui a révélé que la perte de la protéine Atp7a affectait de manière différentielle les cellules épithéliales intestinales, en fonction de leur statut KRAS. De façon intéressante, nous avons constaté que ATP7A était régulé à la hausse dans les cellules avec KRAS muté par rapport à leurs homologues de type sauvage. ATP7A a un double rôle dans les cellules; alors qu'il est essentiel pour la maturation des enzymes dépendantes du cuivre (Cu), ATP7A protège les cellules d'une toxicité excessive induite par le Cu (cuproptose). Chez l'homme, les mutations dans ATP7A entraînent des troubles caractérisés par des déficiences systémiques dans le transport et les niveaux de Cu. Chez les animaux et dans les modèles de culture cellulaire, tel que les cellules épithéliales intestinales, les niveaux intracellulaires de Cu sont directement corrélés avec l'abondance post-transcriptionnelle d'ATP7A. Dans le même ordre d'idées, nous avons observé que les cellules de CCR avec KRAS muté avaient relativement plus de Cu intracellulaire, et la surexpression d'ATP7A protégeait les cellules KRAS muté de la cuproptose, par rapport à leurs homologues de type sauvage. Nous avons également observé que la croissance in vivo des xénogreffes KRAS mutées était réduite lorsque les souris étaient nourries avec un régime pauvre en Cu. Le Cu est utilisé par plusieurs enzymes qui régulent des fonctions cellulaires critiques, notamment la respiration mitochondriale, la motilité cellulaire et la prolifération. Nous montrons que les cellules mutantes KRAS étaient plus sensibles au chélateur de Cu, ammonium tetrathiomolybdate (TTM), par rapport aux cellules de type sauvage. De plus, les cellules avec KRAS muté traitées avec le TTM ont présenté des activités réduites de MEK1/2 dépendant du Cu et de l'enzyme de la chaîne de transport d'électrons mitochondriale, cytochrome c oxidase (CCO). Nous avons été surpris de constater que le transporteur de Cu de haute affinité, CTR1, est régulé à la baisse dans les cellules avec KRAS muté, et avons donc émis l'hypothèse que les cellules KRAS mutées doivent absorber le Cu par d'autres moyens. Ainsi, nous avons constaté que la macropinocytose agit comme une voie non canonique d'approvisionnement en Cu dans les cellules avec KRAS muté. Le traitement de cellules in vivo avec l'inhibiteur de la macropinocytose, EIPA, a inhibé l'expression d'ATP7A et diminué le Cu biodisponible dans les xénogreffes KRAS mutées. En conclusion, nos résultats montrent que les cellules avec KRAS muté augmentent les niveaux de Cu et d'ATP7A pour soutenir la tumorigenèse en augmentant l'activité cuproenzymatique et diminuant la cuproptose. Cette étude est pertinente pour le cancer, car les tissus tumoraux contiennent fréquemment des niveaux de Cu plus élevés que les tissus normaux. Des études récentes ont mis en évidence un potentiel de repositionnement du chélateur de Cu TTM, qui est disponible en clinique et utilisé pour traiter les troubles du Cu. Nos résultats démontrent que la biodisponibilité du Cu pourrait être exploitée pour traiter le CCR avec KRAS muté avec de tels inhibiteurs. Les travaux futurs comprennent l'identification de stratégies combinatoires qui peuvent être améliorer les effets anti-cancéreux de la chélation du Cu. / KRAS is amongst the most frequently mutated genes driving human cancers, including ~ 45% of colorectal cancers (CRC). Despite intense efforts to curb its oncogenic potential, mutant KRAS is frequently associated with drug resistance and is extremely challenging to target therapeutically. Cell-surface proteins are often spatially dysregulated in cancers and are attractive therapeutic targets due to their easy accessibility. We performed RNA sequencing of mutant KRAS-expressing intestinal epithelial cells and observed that cells undergoing transformation exhibited dramatic changes in cell surface-coding genes. Therefore, our goal was to identify novel druggable targets expressed at the cell surface of mutant KRAS-transformed cells. Using a cutting-edge cell surface proteomics approach, we identified several differentially expressed proteins at the surface of KRAS-mutant cells compared to wild-type counterparts. We then performed a cell surface based CRISPR/Cas9 screen, which revealed that loss of the copper exporter Atp7a differentially affected the fitness of intestinal epithelial cells, depending on their KRAS status. Interestingly, we found that ATP7A was upregulated in KRAS-mutant cells compared to wild-type counterparts. ATP7A has a dual role in cells; while it is essential for maturation of copper (Cu)-dependent enzymes, ATP7A protects cells from excess Cu-induced toxicity (cuproptosis). In humans, ATP7A mutations result in disorders characterized by systemic deficiencies in Cu transport and levels. In animals and in tissue culture models, including intestinal epithelial cells, intracellular Cu levels are directly correlated with the post-transcriptional abundance of ATP7A. In line with this, we observed that KRAS-mutant CRC cells and tissues had relatively more intracellular Cu, and ATP7A-overexpression protected KRAS-mutant cells from cuproptosis, compared to wild-type counterparts. We also observed that in vivo growth of KRAS-mutant xenografts was reduced when mice were fed a Cu-deficient diet. Cu is utilized by several enzymes that regulate critical cellular functions including mitochondrial respiration, cell motility and proliferation. We show that KRAS-mutant cells were more sensitive to the Cu chelating drug ammonium tetrathiomolybdate (TTM), compared to wild-type cells. Moreover, TTM-treated KRAS-mutant cells displayed reduced activities of Cu-dependent MEK1/2 and mitochondrial electron transport chain enzyme, cytochrome c oxidase (CCO). We were surprised to find that the high-affinity CTR1 importer is downregulated in KRAS-mutant cells, and so we hypothesized that KRAS cells must uptake Cu through alternate means. In accordance with this, we found that macropinocytosis acts as a non-canonical Cu-supply route in KRAS-mutant cells. In vivo, treating cells with the macropinocytosis inhibitor EIPA, inhibited the expression of ATP7A and decreased bioavailable Cu in KRAS xenografts. In conclusion, our results show that KRAS-mutant cells increase Cu and ATP7A levels, likely to support tumorigenesis by elevating cuproenzymatic activity and parallelly dealing with cuproptosis. This study is relevant to cancer as tumor tissues and patients contain higher Cu levels than normal controls. Recent studies have highlighted a potential for repurposing the clinically available copper chelator TTM, which is used to treat Cu disorders. Our results demonstrate that copper bioavailability could be exploited to treat KRAS-mutated CRC with such inhibitors. Future work includes identification of combinatorial strategies that may be synthetic lethal to copper chelation.

Context: Colorectal cancer (CRC) surgical treatment delay (TD) has been associated with mortality and morbidity; however, disparities by TD profiles are unknown. Objectives: This study aimed to identify CRC patient profiles of surgical TD while accounting for differences in sociodemographic, health insurance, and geographic characteristics. Design: We used latent class analysis (LCA) on 2005-2015 Tennessee Cancer Registry data of CRC patients and observed indicators that included sex/gender, age at diagnosis, marital status (single/married/divorced/widowed), race (White/Black/other), health insurance type, and geographic residence (non-Appalachian/Appalachian). Setting: The state of Tennessee in the United States that included both Appalachian and non-Appalachian counties. Participants: Adult (18 years or older) CRC patients (N = 35 412) who were diagnosed and surgically treated for in situ (n = 1286) and malignant CRC (n = 34 126). Main Outcome Measure: The distal outcome of TD was categorized as 30 days or less and more than 30 days from diagnosis to surgical treatment. Results: Our LCA identified a 4-class solution and a 3-class solution for in situ and malignant profiles, respectively. The highest in situ CRC patient risk profile was female, White, aged 75 to 84 years, widowed, and used public health insurance when compared with respective profiles. The highest malignant CRC patient risk profile was male, Black, both single/never married and divorced/separated, resided in non-Appalachian county, and used public health insurance when compared with respective profiles. The highest risk profiles of in situ and malignant patients had a TD likelihood of 19.3% and 29.4%, respectively. Conclusions: While our findings are not meant for diagnostic purposes, we found that Blacks had lower TD with in situ CRC. The opposite was found in the malignant profiles where Blacks had the highest TD. Although TD is not a definitive marker of survival, we observed that non-Appalachian underserved/underrepresented groups were overrepresented in the highest TD profiles. The observed disparities could be indicative of intervenable risk.

No description available.

Indiana University-Purdue University Indianapolis (IUPUI) / Men’s adherence to masculinity norms has been implicated as a risk factor for unhealthy behaviors (e.g., drinking to intoxication, having unprotected sex with multiple, simultaneous partners) and lack of engagement in healthy behaviors (e.g., blood pressure screening, cholesterol screening, wearing protective clothing while in the sun, receipt of annual medical and dental exams) (Boman & Walker, 2010; Courtenay, 2000a, 2000b, 2011; Hammond, Matthews, & Corbie-Smith, 2010; Iwamoto, Cheng, Lee, Takamatsu, & Gordon, 2011; Locke & Mahalik, 2005; Mahalik, Lagan, & Morrison, 2006; Mahalik et al., 2003; Nicholas, 2000; Pachankis, Westmaas, & Dougherty, 2011; Pleck, Sonenstein, & Ku, 1993; Wade, 2009). Masculinity has been defined as behaviors, beliefs, and personality characteristics associated more often with men than women as well as characteristics and behaviors that society prescribes and reinforces in men (Thompson, Pleck, & Ferrera, 1992). Rooted in geographical, cultural, and temporal environments, diverse masculinities have emerged throughout the United States and the world (Connell, 1995; Courtenay, 2011). Traditional masculinity beliefs and behaviors in the United States include the sturdy oak (men should be tough, self-reliant, stoic, and confident), no sissy stuff (men should avoid feminine characteristics and behaviors), the big wheel (men should strive for success and status), and give ‘em hell (men should embrace aggressiveness, daring, and violence) (Brannon, 1976). Numerous qualitative studies have suggested that some men find cancer screening examinations involving the rectum (i.e., endoscopy for colorectal cancer [CRC] screening or digital rectal examination [DRE] for prostate cancer screening) an affront to their masculinity (see Table 1 for quotations from these studies) (Bass et al., 2011; Beeker, Kraft, Southwell, & Jorgensen, 2000; Getrich et al., 2012; Goldman, Diaz, & Kim, 2009; Harvey & Alston, 2011; Holt et al., 2009; Jilcott Pitts et al., 2013; Jones, Devers, Kuzel, & Woolf, 2010; Rivera-Ramos & Buki, 2011; Thompson, Reeder, & Abel, 2011; Wackerbarth, Peters, & Haist, 2005; Winterich et al., 2009). However, to the author’s knowledge, no quantitative studies have considered the role of masculinity in CRC screening adherence. Unfortunately, current CRC screening rates fall below the 70.5% Healthy People 2020 screening objective (U.S. Department of Health and Human Services, 2012).Research is needed to better understand relationships between men’s masculinity norms and CRC screening adherence so that interventions may be developed to reduce barriers to screening, improve screening rates, and, ultimately, decrease men’s mortality from CRC. The present study will address this gap in the literature by examining the masculinity norms and CRC screening adherence of male veterans aged 51-75 years who are at average CRC risk (Levin et al., 2008). First, the prevalence of CRC, its risk factors and warning signs as well as CRC screening techniques, screening rates, and characteristics of individuals who are adherent and non-adherent to CRC screening guidelines are summarized. Next, the concept of masculinity, theoretical and empirical support for studying masculinity norms within the context of CRC screening, and potential relationships between masculinity norms and colorectal cancer screening behaviors are described. Finally, the study methods, results, and future directions and limitations of this research are described.

Cancers represent second the most common cause of death in the Czech Republic. The most common are breast and colorectal cancers. Identification of prognostic factors improving decision-making approaches for treatment optimization belongs to the key aims of clinical research in oncology. Carriers of mutation in cancer-susceptibility genes represent a small but clinically important group of high-risk patients. The implementation of NGS have accelerated predisposing genes analyses. The large extent of data about the presence of variants in predisposing genes is in striking contrast to only a very limited information available about clinico-pathological characteristics of mutation carriers. Determination of the risk of tumor development in carriers of rare mutations or variants of unclear significance in genes with incomplete penetrance represent substantial drawbacks of current NGS analyses. To address these issues, we have attempted i) to introduce a unified approach to NGS analysis in breast cancer patients, ii) to characterize importance of prognostic factors in BRCA1/BRCA2 mutation carriers, and iii) to identify the cancer risks in carriers of germline mutations in the CHEK2 gene. Colorectal cancer represents seemingly histologically homogeneous disease. However, at the molecular level it can be...

Tjock-och ändtarmscancer (kolorektalcancer) är den tredje vanligaste cancertypen och den näst vanligaste cancer-relaterade dödsorsaken i världen. Östrogen har visat sig ha en skyddande roll mot kolorektalcancer och östrogenreceptor beta är den dominerande östrogenreceptorn i normalt kolonepitel. Immunceller påverkar utvecklingen hos tumörer och forskning på samspelet mellan cancerceller och immunceller kan vara viktig för framtida cancerforskning. Den här studien har för avsikt att undersöka hur koloncancerceller påverkar makrofager och vice versa genom att utföra samkultursexperiment och analysera genuttryck med RT-qPCR. Tumör-associerade makrofager (TAM) polariserades från THP-1 celler och odlades tillsammans med SW480-kolorektalceller med eller utan uttryck av ERβ. En immunfluorescens-analys gjordes på en musmodell av kolit (tarminflammation) för att undersöka andelen av olika immunceller i tjocktarmen. Analysen gjordes i QuPath och beräkningarna mellan en erfaren och en oerfaren användare jämfördes. Denna studie visade att bildandet av TAM:s med hjälp av konditionerat medium från SW480-celler ändrade genuttrycket mot en pro-inflammatorisk fenotyp. Samkultur-experimenten uppvisade motstridiga resultat men antyder att genuttrycket för TAM:s ändras av att vara i samkultur med SW480-celler och att genuttrycket för SW480-celler ändras av att vara i samkultur med THP- 1-celler. Vidare visade resultatet från samkulturen att ERβ-uttrycket i SW480 cellerna påverkade deras genuttryck. Immunofluorescence-analysen av mustarmen demonstrerade att immuncellen med den högsta andelen var dendritiska celler medan celltypen med minst andel var cytotoxiska T-celler, som till antalet var ungefär hälften så många som T-hjälparceller. Analysen visade att skillnaden mellan en erfaren och en oerfaren användare var signifikant för två av tio immunceller. Slutsatserna från denna studie var att SW480-celler har en inverkan på genuttrycket av TAM:s och att genuttrycket i SW480 påverkades av att vara i samodling med THP-1-celler polariserade till makrofagliknande celler (av PMA) eller TAM:s (genom konditionerat medium från SW480-celler). Vidare tycks ERβ påverka uttrycket av ICAM1 och IL-1β i SW480 celler under samkultur med makrofagliknande- eller TAM-THP-1 celler. Genom att fortsätta undersöka förhållandet mellan makrofager och kolorektalcancerceller kan forskningen breddas vilket kan leda till nya behandlingsmetoder i framtiden. / Colorectal cancer (CRC) is the third most common type of cancer and the second leading cause of cancer-related mortality in the world. Estrogen has been found to have a protective role in the development of colorectal cancer and estrogen receptor beta is the predominant estrogen receptor in normal colonic epithelium. Immune cells influence tumor progression and research on the crosstalk between cancer cells and immune cells could be important in future therapies. This study aims to investigate how colorectal cancer cells influence macrophages and vice versa by conducting co-culture experiments and analyzing gene expression using RT-qPCR. Tumor-associated macrophages (TAMs) were polarized from THP-1 cells and cultured together with SW480 colorectal cancer cells with or without the expression of ERβ. Immunofluorescence analysis was performed on colon tissue samples from a colitis-induced mice model to investigate the percentage of different immune cells in the colon. The analysis was done in QuPath and the calculations between an inexperienced user and an experienced user were compared to investigate how the results differ. We found that the formation of TAMs using SW480 conditioned media changed gene expression toward a pro-inflammatory phenotype. The co-culture experiments showed conflicting results but suggest the gene expression of TAMs is altered by being cultured with SW480 and that the gene expression of SW480 cells was affected by being cultured with THP-1 cells. Further, the ERβ expression in SW480 cells affected the gene expression of the cells during co-culture with macrophage-like or TAM THP-1 cells. In the immunofluorescence analysis of mouse colon, the immune cell type with the highest abundance was dendritic cells and the lowest seem to be cytotoxic T-cells, which was around half of the number of T-helper cells. There was a significant difference between the analysis of experienced and inexperienced annotators for two out of ten markers. The conclusions from this study were that SW480 cells have an impact on the gene expression of TAMs and that the gene expression in SW480 was influenced by being in co-culture with THP-1 cells polarized into macrophage-like cells (by PMA) or TAMs (by conditioned media from SW480 cells). Further, ERβ impacted the expression of ICAM1 and IL-1β in SW480 cells during co-culture with macrophage-like or TAM THP-1 cells. By further studying the correlation between macrophages and CRC cells, the research can be broadened which can lead to new approaches to CRC therapies in the future.

No description available.

No description available.

EPHB2, an ephrin receptor (EPH) from receptor tyrosine kinase (RTK) family, is one of the substrates for tissue factor (TF) - coagulation factor VIIa (FVIIa) complex and it is cleaved in its ectodomain. EPHB2 cleavage is important for ephrin receptor (EPH) - ephrin ligand (EFN) signaling and cell repulsion. TF has been reported to be overexpressed in different cancer types such as breast and colorectal cancer (CRC). Furthermore, EPHB2 R155C mutation, at the TF/FVIIa-mediated cleavage site, has been identified as one of the somatic mutation sites in human metastatic CRC. Therefore, the aim of the present work was to characterize the EPHB2 R155C mutation and its effect on the cleavage by TF/FVIIa on EPHB2 in context to CRC. We generated overexpression cell models for EPHB2 wild type (wt) and R155C mutant in human CRC DLD-1 cell line for in vitro compartmentalization assay analysis to demonstrate repulsion event in EPH-EFN signaling. Whereas low endogenous TF expression led to incomplete cleavage of EPHB2 wt protein, stable overexpression of TF resulted in complete cleavage. Moreover, overexpression of TF resulted in reduced compartmentalization in EPHB2 wt cells after FVIIa treatment. Transient expression of TF in EPHB2 wt and R155C cells showed no clear difference in EPHB2 cleavage. Interestingly, it was difficult to obtain similar stable overexpression level of TF in EPHB2 R155C cells compared to EPHB2 wt cells. This may lead to further research in context to the role of TF/FVIIa-mediated EPHB2 cleavage in CRC by the generation of TF overexpression cell lines using lentiviral transduction.

Bakgrund: Patienter med kolorektalcancer vårdas enligt enhanced recovery after surgery (ERAS) vårdprogram. ERAS är ett evidensbaserat vårdprogram som syftar till att effektivisera och förbättra vårdkvalitén genom hela den perioperativa vårdprocessen. Den perioperativa dialogen är nyckeln till god vårdrelation inom ERAS för att motverka vårdlidande. Operationssjuksköterskans funktion är betydelsefull i vårdandet av dessa patienter. Syfte: Belysa operationssjuksköterskors erfarenheter av att vårda patienter som genomgår kolorektalkirurgi inom ERAS vårdprogram perioperativt. Metod: En kvalitativ studie där åtta operationssjuksköterskor intervjuades. Insamlade data analyserades med hjälp av en kvalitativ innehållsanalys. Resultat: Informanternas upplevelser av att vårda patienter enligt ERAS vårdprogram var positivt. ERAS vårdprogram ansågs vara strukturerat och effektivt. ERAS resulterade i total optimering hos patienter som ska genomgå kolorektalkirurgi. Flera fördelar kunde identifieras med ERAS vårdprogrammet dock framkom det att operationssjuksköterskornas delaktighet var minimal. Däremot hade operationsteamets samverkan en central betydelse i arbetet med ERAS vårdprogram. Konklusion: Arbetet och resultaten inom kirurgisk vård har förbättrats avsevärt efter implementeringen av ERAS. ERAS skapar goda förutsättningar för patienterna i deras vård. / Background: Patients with colorectal cancer gets treatment within the ERAS program. It is an evidence-based program with the purpose of making the care more effective through the perioperative process. The perioperative dialogue is the key to a good care relationship within the ERAS to counteract care suffering. The role of the operating room nurse is significant in the care of these patients. Aim: The perioperative experience from operating room nurses when they are taking care of patients with colorectal cancer within the ERAS program. Method: A quality based study with interviews from eight operating room nurses. Collected data which was analyzed with the help of a quality based content analysis.  Result: the experience of taking care of patients within ERAS were positive by the informants. ERAS is considered being a structured and effective program. It resulted in an optimization with patients undergoing colorectal surgical. A lot of positive things were identified within ERAS, although the participation of operating room nurses were minimal. However were the cooperation of the surgical team important within the ERAS program.  Conclusion: The work and result within surgical care has improved significantly since the introduction of ERAS. The program creates good conditions for patients in healthcare.