Multiplexed cellular profiling identifies an organoselenium compound as an inhibitor of CRM1-mediated nuclear export.

Chromosomal region maintenance 1 (CRM1 also known as Xpo1 and exportin-1) is the receptor for the nuclear export controlling the intracellular localization and function of many cellular and viral proteins that play a crucial role in viral infections and cancer. The inhibition of CRM1 has emerged as a promising therapeutic approach to interfere with the lifecycle of many viruses, for the treatment of cancer, and to overcome therapy resistance. Recently, selinexor has been approved as the first CRM1 inhibitor for the treatment of multiple myeloma, providing proof of concept for this therapeutic option with a new mode of action. However, selinexor is associated with dose-limiting toxicity and hence, the discovery of alternative small molecule leads that could be developed as less toxic anticancer and antiviral therapeutics will have a significant impact in the clinic. Here, we report a CRM1 inhibitor discovery platform. The development of this platform includes reporter cell lines that monitor CRM1 activity by using red fluorescent protein or green fluorescent protein-labeled HIV-1 Rev protein with a strong heterologous nuclear export signal. Simultaneously, the intracellular localization of other proteins, to be interrogated for their capacity to undergo CRM1-mediated export, can be followed by co-culturing stable cell lines expressing fluorescent fusion proteins. We used this platform to interrogate the mode of nuclear export of several proteins, including PDK1, p110α, STAT5A, FOXO1, 3, 4 and TRIB2, and to screen a compound collection. We show that while p110α partially relies on CRM1-dependent nuclear export, TRIB2 is exported from the nucleus in a CRM1-independent manner. Compound screening revealed the striking activity of an organoselenium compound on the CRM1 nuclear export receptor.

mTORC2 Is the Major Second Layer Kinase Negatively Regulating FOXO3 Activity.

Forkhead box O (FOXO) proteins are transcription factors involved in cancer and aging and their pharmacological manipulation could be beneficial for the treatment of cancer and healthy aging. FOXO proteins are mainly regulated by post-translational modifications including phosphorylation, acetylation and ubiquitination. As these modifications are reversible, activation and inactivation of FOXO factors is attainable through pharmacological treatment. One major regulatory input of FOXO signaling is mediated by protein kinases. Here, we use specific inhibitors against different kinases including PI3K, mTOR, MEK and ALK, and other receptor tyrosine kinases (RTKs) to determine their effect on FOXO3 activity. While we show that inhibition of PI3K efficiently drives FOXO3 into the cell nucleus, the dual PI3K/mTOR inhibitors dactolisib and PI-103 induce nuclear FOXO translocation more potently than the PI3Kδ inhibitor idelalisib. Furthermore, specific inhibition of mTOR kinase activity affecting both mTORC1 and mTORC2 potently induced nuclear translocation of FOXO3, while rapamycin, which specifically inhibits the mTORC1, failed to affect FOXO3. Interestingly, inhibition of the MAPK pathway had no effect on the localization of FOXO3 and upstream RTK inhibition only weakly induced nuclear FOXO3. We also measured the effect of the test compounds on the phosphorylation status of AKT, FOXO3 and ERK, on FOXO-dependent transcriptional activity and on the subcellular localization of other FOXO isoforms. We conclude that mTORC2 is the most important second layer kinase negatively regulating FOXO activity.

P057 How Distance Between Residence and Treatment Center Impact the Outcome of Patients With Inflammatory Bowel Disease at CMN 20.

BACKGROUND: Inflammatory bowel disease (IBD) affects an estimated 1.6 million people in the United States with at least 70,000 new cases per year. In the last 15 years an increase in frequency of IBD cases from 0.30 cases per 100,000 person-years to 1.83 cases per 100,000 person-years has been reported in Mexico. Variables such as early hospitalization at diagnosis, low hemoglobin levels, high values of C-reactive protein (CRP), fecal calprotectin concentration and absence of mucosal healing during evolution define the prognosis of these patients. However, the distance a patient must travel in order to reach a specialized treatment center can become an impediment for a correct diagnosis and treatment, severely impacting the clinical outcome of such patients. METHODS: Observational, cross-sectional, retrospective study. Objective: In patients with IBD, determine the impact of distance between the residence and specialized treatment center on the general clinical outcome. Variables analyzed: sex, age, disease duration, average travel time, distance in kilometers between place of residence and specialized treatment center, Crohn´s Disease (CD) or Ulcerative Colitis (UC), and their respective severity classification scores, as well as number of hospital readmissions per year. The results were evaluated with ANOVA tests, univariate analysis had a 95% confidence index and a significant "p" determined as p < 0.05. RESULTS: The study included 66 patients (45 UC and 21 CD). Mean age 51.15± 17.5 years. The distance between residence and hospital was calculated and classified into 3 quartiles based on proximity: 750km (quartile 3). There was a higher risk among patients in the most distant quartile for the use of biological therapy (OR, 2.20; 95% CI, 0.23- 20.55) and surgery (OR, 2.76, 95% CI 0.49- 15.48). We observed a clear relationship between the number of hospital readmission and the distance quartiles with a p = 0.0047. CONCLUSION: We observed an impact between the distance of residence and specialized treatment center over the patient's clinical outcome. More readmissions, greater disease activity scores, more use of biological therapy and surgery were observed in patients who had to travel more than 750km from their home to their specialized treatment center compared to the other travel quartiles, this with a p = 0.0047.

P061 Impact of Inflammatory Bowel Disease on sleep quality in a Mexican population attended in a referral center.

BACKGROUND: Sleep disorders occur recurrently in patients with inflammatory bowel disease (IBD). The relationship between poor sleep quality and IBD activity has been subject to scarce attention. Poor sleep quality could be considered a relevant extraintestinal manifestation and a potential marker of subclinical inflammation, which could increase the severity of inflammation and the risk of relapse, however, we do not have enough information to confirm this hypothesis. Objective: Describe the impact of IBD on the quality of sleep, in patients treated in a referral hospital. METHODS: Observational, analytical, and cross-sectional study. Patients with diagnosis of IBD treated at Centro Médico Nacional "20 de Noviembre" were evaluated. The Pittsburgh Sleep Quality Index (PSQI) was used to measure sleep quality. IBD activity was measured using the Harvey-Bradshaw index for Crohn's disease (CD) and the Mayo scale for Ulcerative Colitis (UC). RESULTS: A total of 51 patients were included, the Pittsburgh Sleep Index Questionnaire (PSQI) was performed, after informed consent was signed. Patients had a mean age of 52. Fifty five percent were female, 65% had UC. Biological therapy was administered to 70.5%. In CD 89% were in remission and 11% in moderate activity. In UC 48.5% were in remission, 45.5% had mild activity and 6% had moderate activity. Eighty percent of the patients did not use hypnotic drugs. Patients with UC in remission had a bad perception of sleep quality in 68%, quite good sleep quality in 18% and very good sleep quality in 12% with a PSQI of 10.5 ± 3.2. In patients with mild activity, the perception of sleep quality was very good in 6%, quite good in 46%, quite bad in 40% and very bad in 6%, with a PSQI of 8 ± 3.7. In patients with moderate activity, 100% had a rather bad perception of sleep quality with a PSQI of 11 ± 1.4. For CD in remission the perception of sleep was quite bad in 43%, quite good in 43%, very bad in 6% and very good in 6% with a PSQI of 9 ± 4.3. In patients with moderate activity 50% had a very bad sleep quality perception and 50% a fairly good sleep quality perception with a PSQI of 14 ± 4.2. CONCLUSION: In this study a statistically significant association was obtained between PSQI and the perception of sleep reported by the patients, with a p < 0.005. Further research is still needed to better characterize sleep disturbances in this population. Due to the sample size, a prospective, randomized study is required to confirm these findings. The present analysis has no conflict of interest.

P063 Prevalence in the Use of Complementary/Alternative Medicine in Patients With Inflammatory Bowel Disease from Centro Médico Nacional 20 de Noviembre.

BACKGROUND: Frequently, existing therapies for inflammatory bowel disease (IBD) do not achieve complete remission of the disease and are commonly associated with adverse effects. Therefore, patients regularly turn to complementary or alternative medicine (CAM). International CAM use in IBD patients ranges from 21% to 60%. There are no reported statistics of CAM use in the Mexican population who suffer IBD. This study´s objective was to determine the prevalence and characterize the use of CAM in our IBD population. METHODS: Observational, cross-sectional, retrospective study in patients diagnosed with IBD belonging to our Institution's IBD center. The results were analyzed with measures of relative frequency, central tendency, and dispersion. RESULTS: A total of 52 patients with IBD (78% ulcerative colitis and 22% Crohn's disease) were included, 38.5% were exposed to CAM. A total of 27% used it as an aid in their IBD treatment. The remaining 11.5% used it for other reasons. Patients exposed to CAM for IBD averaged 51 ± 19 years, 64% were female and 71% had at least an undergraduate degree. The most used CAMs were acupuncture (42.8%), herbal (35.7%) and homeopathy (35.7%). Half of the patients used concomitantly two or more modalities, and 91.6% of the patients knew their diagnosis at the time that CAM was being used. The median exposure time to CAM was 4.5 ± 40.7 months. The main reasons for the use of CAM in IBD patients was as a complementary therapy in 58.3%, as the perception of lack of improvement was present in 33.3% of patients. Twenty-five percent of patients used CAM after a healthcare professional recommended it. CAM was used in 83.2% of patients with conventional treatment (Mesalamine: 74.6%, Mesalamine + Azathioprine: 8.3%) and only 16.5% were on biological therapy. A total of 58.3% CAM users perceived improvement in their symptoms. Currently 25% still use some modality of CAM. No statistically significant differences were found in sociodemographic variables and clinical outcomes when comparing the IBD group exposed to CAM vs the unexposed group. CONCLUSION: Exposure to CAM for IBD treatment had a prevalence of 27%. The main CAM modalities for IBD patients were acupuncture, herbal, and homeopathy. No relationship was demonstrated between the use of CAM in IBD patients and their clinical outcomes.

P073 Diagnostic Outcome of Patients With Non-Specific Colitis.

BACKGROUND: Colitis describes inflammation in the colon and is classified into defined diseases: Inflammatory bowel disease (IBD), Microscopic colitis, Ischemic colitis and Infectious colitis. The clinical significance of non-specific colitis is unclear as there are limited analysis which report the issue. Notteghem et al. (1993) showed that of 104 patients with non-specific colitis, 52.3% had another episode of symptoms within 3 years. Of these patients 54% were diagnosed with ulcerative colitis, 33% with Crohn's disease, and 13% remained unclassified, suggesting that non-specific colitis could be undiagnosed IBD. The objective of this analysis is to determine the clinical course of non-specific colitis through colonoscopy and histologic findings. METHODS: Retrospective observational study with patients underwent colonoscopies during the period from 2009 to 2019, whose biopsies were reported as non-specific colitis. RESULTS: 24 patients (18 women and 6 men) with non-specific colitis undergoing follow-up by the Gastroenterology outpatient clinic were included. The most frequent indications for colonoscopy were abdominal pain (43.7%), blood in stools (26.8%), diarrhea (21.6%) and anemia (7.9%). The mean age was 56.5 years. During follow-up, patients had at least 2 colonoscopies and 4 biopsies per colonic segment. In the male population ascending colon cancer was later diagnosed in 16.7%, another 16.7% were diagnosed with nonspecific chronic ulcerative colitis, 33.3% with eosinophilic colitis, and the rest of the patients were diagnosed with non-specific colitis. From the female group, 5.6% had lymphocytic colitis, 5.6% eosinophilic colitis, 5.6% collagenous colitis, 5.6% Crohn's disease, 5.6% mixed irritable bowel syndrome, and the rest of the patients (72%) had biopsy report of non-specific colitis. CONCLUSIONS: All patients with a histopathological diagnosis of non-specific colitis underwent a second colonoscopy with biopsy, which resulted in a definitive diagnosis of Crohn's disease in 5.6% of women and Ulcerative Colitis in 16.7% of men. This study comes to show that there is an important subgroup of patients who are underdiagnosed. We suggest that patients with non-specific colitis reported on a histopathology report must continue their follow-up by a specialized ward.

[Iatrogenic ureteral injury]. / Trauma de uréter yatrógeno.

OBJECTIVES: To determine the most frequent causes of ureteral injury, and the way to correct severe damage of the ureter. METHODS: We reviewed the medical records of 25 patients who suffered ureteral injuries during abdominal or pelvic surgery in a five-year period; endoscopic injuries were excluded. RESULTS: The most common presentation included pain, urinary incontinence and fever. Hysterectomy was the most frequent operation causing ureteral injuries. The diagnosis was made by intravenous urography and ultrasound. Ureteral reimplantation using a Boari's flap, without other complications, was the preferred treatment due to ureteral damage. Right side and distal third of the ureter where the most frequently affected sites. CONCLUSIONS: In cases of iatrogenic injuries of the distal third of the ureter with severe lesions, ureteral reimplantation with a Boari's flap may be the option.