Search

Supplemental Methods and Materials. ACF Phospho-ERK and ERK Level Quantification.

Colorectal cancer progresses through multiple distinct stages that are potentially amenable to chemopreventative intervention. Epidermal growth factor receptor (EGFR) inhibitors are efficacious in advanced tumors including colorectal cancer. There is significant evidence that EGFR also plays important roles in colorectal cancer initiation, and that EGFR inhibitors block tumorigenesis. We performed a double-blind randomized clinical trial to test whether the EGFR inhibitor erlotinib given for up to 30 days had an acceptable safety and efficacy profile to reduce EGFR signaling biomarkers in colorectal aberrant crypt foci (ACF), a subset of which progress to colorectal cancer, and normal rectal tissue. A total of 45 patients were randomized to one of three erlotinib doses (25, 50, and 100 mg) with randomization stratified by nonsteroidal anti-inflammatory drug (NSAID) use. There were no unanticipated adverse events with erlotinib therapy. Erlotinib was detected in both normal rectal mucosa and ACFs. Colorectal ACF phosphorylated ERK (pERK), phosphorylated EGFR (pEGFR), and total EGFR signaling changes from baseline were modest and there was no dose response. Overall, this trial did not meet is primary efficacy endpoint. Colorectal EGFR signaling inhibition by erlotinib is therefore likely insufficient to merit further studies without additional prescreening stratification or potentially longer duration of use. Cancer Prev Res; 8(3); 222–30. ©2015 AACR.

Perspective on this Article from Risk of Cardiovascular Events in a Randomized Placebo-Controlled, Double-Blind Trial of Difluoromethylornithine plus Sulindac for the Prevention of Sporadic Colorectal Adenomas

On the basis of large cardiovascular clinical trials of lipid-lowering agents that showed a considerable decrease in the incidence of primary melanomas in the active agent arm, we have carried out a randomized, double-blind clinical trial examining the impact of lovastatin on various biomarkers of melanoma pathogenesis. Subjects with at least two clinically atypical nevi were randomized to receive oral lovastatin or placebo for a 6-month period. Clinical, histopathologic, and molecular biomarkers were evaluated for change in the two groups. Eighty subjects were randomized, evaluable, and included in the analyses. Lovastatin showed no benefit in comparison with placebo in the primary endpoint of decreasing the level of histopathologic atypia, nor in any of the secondary endpoints of decreasing clinical atypia, impact on nevus number, nor in showing significant changes in any of the molecular biomarkers. There were no significant differences in adverse event profiles for lovastatin compared with placebo. The lovastatin arm did show a significant and considerable decrease in total serum cholesterol and serum low-density lipoprotein (LDL) levels compared with placebo, an expected result. This finding bolsters confidence in subject compliance. Given the results of this trial, it is concluded that if lovastatin were to lower the incidence of melanoma, it would appear not to be doing so by reversing atypia of precursor atypical nevi over the 6-month time frame studied. Further research into the pathogenesis of melanoma and in other potential chemopreventive agents is needed. Cancer Prev Res; 7(5); 496–504. ©2014 AACR.

Supplemental Figure 1. Metformin effects on insulin, AMPK, mTOR

legend for supplementary figure 1

PDF file - 3911KB, Supplementary Figure S1. Examples of classifications of nevomelanocytic neoplasms histopathologically evaluated for the primary endpoint of the study. Supplementary Figure S2. Examples of a target atypical nevus photographed pre and post treatment. Supplementary Figure S3. Examples of back photos pre and post treatment. Supplementary Figure S4. CONSORT Flowchart for all study subjects. Supplementary Figure S5. Bland-Altman plot of histopathologic regression of atypical nevi. Supplementary Figure S6. Assessment of clinical regression of atypia. Supplementary Figure S7. Bland-Altman plots of (A) HIF-1 percent of nuclear staining, (B) e-Cadherin percent of cytoplasmic staining, (C) N-Cadherin percent of cytoplasmic staining and (D) VEGF percent of cytoplasmic staining. Supplementary Figure S8. Bland-Altman plots of (A) RelA percent of cytoplasmic staining, (B) p21 percent of nuclear staining, and (C) ki-67 percent of nuclear staining.

In animal models of hepatocellular carcinoma (HCC), deficiency of S-adenosylmethionine (SAMe) increased the risk of HCC whereas administration of SAMe reduced HCC. The aim of this trial was to determine whether oral SAMe administration to patients with hepatitis C cirrhosis would decrease serum α-fetoprotein (AFP) level, a biomarker of HCC risk in hepatitis C. This was a prospective, randomized, placebo-controlled, double-blind trial of SAMe, up to 2.4 g/d, for 24 weeks as compared with placebo among subjects with hepatitis C cirrhosis and a mildly elevated serum AFP. Primary outcome was change in AFP between baseline and week 24. Secondary outcomes included changes in routine tests of liver function and injury, other biomarkers of HCC risk, SAMe metabolites, markers of oxidative stress, and quality of life. One hundred ten subjects were randomized and 87 (44 SAMe and 43 placebo) completed treatment. There was no difference in the change in AFP during 24 weeks among subjects receiving SAMe as compared with placebo. Changes in markers of liver function, liver injury, and hepatitis C viral level were not significantly different between groups. Similarly, SAMe did not change markers of oxidative stress or serum glutathione level. SAMe blood level increased significantly among subjects receiving SAMe. Changes in quality of life did not differ between groups. Overall, this trial did not find that SAMe treatment improved serum AFP in subjects with advanced hepatitis C cirrhosis and a mildly elevated AFP. SAMe did not improve tests of liver function or injury or markers of oxidative stress or antioxidant potential. Cancer Prev Res; 8(9); 864–72. ©2015 AACR.

PDF file - 53KB

Perspective on this Article from Longitudinal Assessment of Air Conduction Audiograms in a Phase III Clinical Trial of Difluoromethylornithine and Sulindac for Prevention of Sporadic Colorectal Adenomas

Supplemental Table 1: Change from Week 0 to Week 24 for routine blood tests of liver function or injury. Supplemental Table 2: Compliance with study medicine among 87 subjects who completed the study. Supplemental Table 3: Change from Week 0 to Week 24 in quality of life as assessed with SF-36 and CLDQ.

PDF file - 111KB

Supplemental Figure 1a: SF-36. SF-36 mental component score among subjects randomized to SAMe or placebo, at baseline, Week 12 and Week 24. Supplemental Figure 1b: SF-36 physical component score among subjects randomized to SAMe or placebo, at baseline, Week 12 and Week 24. Only subjects with SF-36 at all three time points were included in the analysis. Supplemental Figure 2: CLDQ. Chronic liver disease questionnaire among subjects randomized to SAMe or placebo, at baseline, Week 12 and Week 24. Only subjects with CLDQ at all three time points were included in the analysis.

PDF file - 64KB, Supplementary Table S1. Descriptive statistics for concordance evaluation of positive control slides. Supplementary Table S2. Evaluation of laboratory data.

Background Fractures are a great health issue associated with morbidity, quality of life, life span, and health care expenditure. Fractures are correlated with cardiovascular disease, type 2 diabetes mellitus, cerebrovascular disease, and some psychiatric disorders. However, representative national data are few, and longitudinal cohort studies on the association between schizophrenia and the subsequent fracture risk are scant. We designed a nationwide population-based cohort study to investigate the association of schizophrenia with hip, vertebral, and wrist fractures over a 10-year follow-up. Methods Data of patients with schizophrenia (International Classification of Diseases, Ninth Revision, Clinical Modification code 295) and matched over January 2000–December 2009) were extracted from Taiwan National Health Insurance Research Database. A Cox proportional-hazards regression model was constructed to calculate hazard ratios (HRs) for fractures between the schizophrenia and control cohorts. Results Of 2028 people with schizophrenia (mean age: 36.3 years, 49.4% female), 89 (4.4%) reported newly diagnosed fractures—significantly higher than the proportion in the control population (257, 3.2%; P = 0.007). The incidences of hip (1.2%, P = 0.009) and vertebral (2.6%, P = 0.011) fractures were significantly higher in the schizophrenia cohort than in the control cohort. In Cox regression analysis, hip (adjusted HR: 1.78, 95% confidence interval [CI]: 1.08–2.93) and vertebral (adjusted HR: 1.40, 95% CI: 1.01–1.95) fracture risks were significantly higher in patients with schizophrenia. Furthermore, a sex-based subgroup analysis revealed that the risk of hip fracture remained significantly higher in female patients with schizophrenia (HR: 2.68, 95% CI: 1.32–5.44) than in female controls. On the other hand, there was no significant interaction between effects of sex and schizophrenia on the risk of fractures. Conclusions Over a 10-year follow-up, hip and vertebral fracture risks were higher in the people with schizophrenia than in the controls. The risk of fractures in patients with schizophrenia does not differ between female and male.

Hoyeraal-Hreidarsson syndrome (HHS) is the most severe form of dyskeratosis congenita (DC) and is caused by genes involved in telomere maintenance. Here, we identified male siblings from a family with HHS carrying a hemizygous mutation (c.1345C>G, p.R449G), located in the C-terminal nuclear localization signal (NLS) of the DKC1 gene. These patients exhibit progressive cerebellar hypoplasia, recurrent infections, pancytopenia due to bone marrow failure, and short leukocyte telomere lengths. Single-cell RNA sequencing analysis suggested defects in the NLRP3 inflammasome in monocytes and the activation and maturation of NK cells and B cells. In experiments using induced pluripotent stem cells (iPSCs) from patients, DKC1_R449G iPSCs had short telomere lengths due to reduced levels of human telomerase RNA (hTR) and increased cytosolic proportions of DKC1. Treatment with dihydroquinolizinone RG7834 and cordycepin rescued telomere length in patient-derived iPSCs. Together, our findings not only provide new insights into immunodeficiency in DC patients but also provide treatment options for telomerase insufficiency disorders.

Understanding how excitable cells work in health and disease and how that behavior can be altered by small molecules or genetic manipulation is important. Genetically encoded calcium indicators (GECIs) with multiple emission windows can be combined (e.g., for simultaneous observation of distinct subcellular events) or used in extended applications with other light-dependent actuators in excitable cells (e.g., combining genetically encoded optogenetic control with spectrally compatible calcium indicators). Such approaches have been used in primary or stem cell-derived neurons, cardiomyocytes, and pancreatic beta-cells. However, it has been challenging to increase the throughput, or duration of observation, of such approaches due to limitations of the instruments, analysis software, indicator performance, and gene delivery efficiency. Here, a high-performance green GECI, mNeonGreen-GECO (mNG-GECO), and red-shifted GECI, K-GECO, is combined with optogenetic control to achieve all-optical control and visualization of cellular activity in a high-throughput imaging format using a High-Content Imaging System. Applications demonstrating cardiotoxicity testing and phenotypic drug screening with healthy and patient-derived iPSC-CMs are shown. In addition, multi-parametric assessments using combinations of spectral and calcium affinity indicator variants (NIR-GECO, LAR-GECO, and mtGCEPIA or Orai1-G-GECO) are restricted to different cellular compartments are also demonstrated in the iPSC-CM model.

569 Background: CPIs targeting PD-1/PD-L1 and CTLA-4 have transformed management of GU cancers. These agents are associated with a unique type of immune-mediated toxicities that may affect any organ, which may be severe or life-threatening. Development of irAEs may in part depend on type of CPI used, type of solid tumor, and individual patient factors including presence of subclinical inflammatory states. However, distinct biomarkers predictive of irAE development are poorly defined. This study aimed to examine possible associations between tumor mutation burden (TMB) and PD-L1 expression with occurrence of irAEs. Methods: A retrospective chart review was performed of all patients with urothelial carcinoma (UC) and renal cell carcinoma (RCC), who received therapy with CPIs from 1/2019 to 9/2022 at our institution and developed associated irAEs. Severe irAEs were defined as those necessitating initiation of immunosuppressive therapies or other supportive therapies. TMB and PDL-1 expression patterns were analyzed, if available. Given lack of standardized cutoffs for PD-L1 positivity across different GU cancer types, tumors were considered positive if a combined positive score of greater than 10 was present in UC or a total positive score of greater than 1% was present in RCC. Results: A total of 60 patients were included in this analysis - 33 with UC and 27 with RCC. Severe irAEs occurred in 19 patients with UC and 25 patients with RCC. Median TMB was 8.7 in patients with UC, while it was 2.5 in patients with RCC. PD-L1 positivity was seen in 39.3% (13/33) of UC patients and 40.7% (11/27) of RCC patients. In the UC cohort, 57.8% (11/19) had severe irAEs and TMB less than 10, while 36.8% (7/19) had severe irAEs and TMB greater than 10. Further, 47.3% (9/19) of UC patients had severe irAEs and positive PD-L1, while 42.1% (8/19) had severe irAEs and negative PD-L1. In the RCC cohort, 88% (22/25) had severe irAEs and TMB less than 10, while 8% (2/25) had severe irAEs and TMB greater than 10. Additionally, 40% (10/25) RCC patients had severe irAEs and positive PD-L1, while 56% (14/25) had severe irAEs and negative PD-L1. The incidence of TERT promoter mutations and TMB is noted in the table. Conclusions: Identification of predictive biomarkers of irAEs from CPIs is a major area of need in GU malignancies. Definitive conclusions from this dataset are limited by its small sample size. Interestingly, there appear to be differences in baseline expression of traditional immune biomarkers between UC and RCC, with lower prevalence of TMB greater than 10 and positive PD-L1 noted in the RCC cohort. As such, their utility when making treatment decisions in clinical practice may be of limited value. [Table: see text]

Multifocal illumination can improve image acquisition time compared to single point scanning in confocal microscopy. However, due to an increase in the system complexity, obtaining uniform multifocal illumination throughout the field of view with conventional methods is challenging. Here, we propose a volume holographic lenslet array illuminator (VHLAI) for multifocal confocal microscopy. To obtain uniform array illumination, a super Gaussian (SG) beam has been incorporated through VHLAI with an efficiency of 43%, and implemented in a confocal microscope. The design method for a photo-polymer based volume holographic beam shaper is presented and its advantages are thoroughly addressed. The proposed system can significantly improve image acquisition time without sacrificing the quality of the image. The performance of the proposed multifocal confocal microscopy was compared with wide-field images and also evaluated by measuring optically sectioned microscopic images of fluorescence beads, florescence pollen grains, and biological samples. The proposed multifocal confocal system generates images faster without any changes in scanning devices. The present method may find important applications in high-speed multifocal microscopy platforms.

Manipulation and precise delivery of optical energies in the regions of interest within specimens require different strategies. Hence, proper control of input beam parameters is a prerequisite. One of the prominent methods is metasurface optics, capable of crafting properties of light at nanoscales. Here, the generation of an abrupt autofocusing (AAF) beam by a nanophotonic metasurface for biomedical applications is demonstrated. Fluorescence guided laser microprofiling of mouse cardiac samples is experimentally investigated, using the AAF beam to deliver optical energy selectively to specific locations. In addition, photocoagulation of ex vivo swine skin tissue is performed and observed through optical coherence tomography. The results show great potentials for integrating metasurface optics to realize miniature laser surgery instruments for wide applications in biomedicine.

526 Background: The impact of COVID-19 on cancer patients may be attributed not only to its direct effects on the immune system but also to delays in diagnosis and treatment. Data on the effects of COVID-19 on pancreatic ductal adenocarcinoma (PDAC) patients are scarce. Therefore, we set out to determine the impact of the pandemic on diagnosis and treatment initiation. We hypothesized that time from diagnosis to treatment would be increased in the COVID era compared to the pre-COVID era. Methods: We conducted an IRB-approved retrospective chart review of 488 patients diagnosed with PDAC from March 2019 to September 2020 at two academic medical centers. Patients were divided into two groups, based on the date of initial pathologic diagnosis. We defined the pre-COVID era as March 2019 to March 2020, the 12-month time period before California’s statewide lockdown. The COVID era was defined as the 6 months following the lockdown, March 2020 to September 2020. Demographics, clinical stage, and treatment type were recorded. In addition, initial clinical encounter date, pathologic diagnosis date, and initial treatment date were also collected. All data were gathered at two large-scale academic institutions. Descriptive statistics were used in the analysis. Results: There were 333 patients diagnosed during the pre-COVID era and 155 patients during the COVID era. While race/ethnicity and age at diagnosis were statistically similar for both groups, females made up a significantly larger proportion of COVID era patients than pre-COVID era patients ( p= 0.02). There was no significant difference in clinical stage at diagnosis between the two groups ( p= 0.84). In the pre-COVID era, 19.5% of cases were resectable, 11.1% borderline resectable, 20.1% locally advanced, and 31.8% metastatic. In the COVID era, 17.4% of patients were resectable, 11% borderline resectable, 23.9% locally advanced, and 32.9% metastatic. Median time from pathologic diagnosis to initiation of treatment was 32 days for the pre-COVID era patients and 28 days for the COVID era patients ( p= 0.38). Initial treatment type was also similar between the two groups ( p= 0.29). Conclusions: Fortunately, our data indicate that the COVID-19 pandemic has not significantly prevented PDAC patients from seeking care. Additionally, it does not appear that COVID-19 has delayed treatment initiation or changed initial treatment type. We believe that the successful adoption of telemedicine and other safety protocols have allowed patients with PDAC to continue receiving appropriate care during the pandemic.[Table: see text]