ctDNA MRD Detection and Personalized Oncogenomic Analysis in Oligometastatic Colorectal Cancer From Plasma and Urine.

We hypothesized that circulating tumor DNA (ctDNA) molecular residual disease (MRD) analysis without prior mutational knowledge could be performed after neoadjuvant chemotherapy to assess oligometastatic colorectal cancer (CRC) treated surgically with curative intent. We also investigated urine as an alternative analyte for ctDNA MRD detection in this nongenitourinary setting. PATIENTS AND METHODS: We applied AVENIO targeted next-generation sequencing to plasma, tumor, and urine samples acquired on the day of curative-intent surgery from 24 prospectively enrolled patients with oligometastatic CRC. Age-related clonal hematopoiesis was accounted for by removing variants also present in white blood cells. Plasma and urine ctDNA MRD were correlated with tumor cells detected in the surgical specimen, and adjuvant treatment strategies were proposed based on ctDNA-inferred tumor mutational burden (iTMB) and targetable alterations. RESULTS: Seventy-one percent of patients were treated with neoadjuvant chemotherapy. Tumor-naive plasma ctDNA analysis detected MRD at a median level of 0.62% with 95% sensitivity and 100% specificity, and 94% and 77% sensitivity when only considering patients treated with neoadjuvant chemotherapy and putative driver mutations, respectively. In urine, ctDNA MRD detection specificity remained high at 100%, but sensitivity decreased to 64% with median levels being 11-fold lower than in plasma (P < .0001). Personalized ctDNA MRD oncogenomic analysis revealed 81% of patients might have been candidates for adjuvant immunotherapy based on high iTMB or targeted therapy based on actionable PIK3CA mutations. CONCLUSION: Tumor-naive plasma ctDNA analysis can sensitively and specifically detect MRD in patients with oligometastatic CRC after neoadjuvant chemotherapy. Urine-based ctDNA MRD detection is also feasible; however, it is less sensitive than plasma because of significantly lower levels. Oligometastatic patients with detectable MRD may benefit from additional personalized treatment based on ctDNA-derived oncogenomic profiling.

Endoplasmic reticulum stress controls M2 macrophage differentiation and foam cell formation.

Macrophages are essential in atherosclerosis progression, but regulation of the M1 versus M2 phenotype and their role in cholesterol deposition are unclear. We demonstrate that endoplasmic reticulum (ER) stress is a key regulator of macrophage differentiation and cholesterol deposition. Macrophages from diabetic patients were classically or alternatively stimulated and then exposed to oxidized LDL. Alternative stimulation into M2 macrophages lead to increased foam cell formation by inducing scavenger receptor CD36 and SR-A1 expression. ER stress induced by alternative stimulation was necessary to generate the M2 phenotype through JNK activation and increased PPARγ expression. The absence of CD36 or SR-A1 signaling independently of modified cholesterol uptake decreased ER stress and prevented the M2 differentiation typically induced by alternative stimulation. Moreover, suppression of ER stress shifted differentiated M2 macrophages toward an M1 phenotype and subsequently suppressed foam cell formation by increasing HDL- and apoA-1-induced cholesterol efflux indicating suppression of macrophage ER stress as a potential therapy for atherosclerosis.

1,25(OH)2 vitamin d inhibits foam cell formation and suppresses macrophage cholesterol uptake in patients with type 2 diabetes mellitus.

BACKGROUND: Cardiovascular disease is the leading cause of death among those with diabetes mellitus. Vitamin D deficiency is associated with an increased risk of cardiovascular disease in this population. To determine the mechanism by which vitamin D deficiency mediates accelerated cardiovascular disease in patients with diabetes mellitus, we investigated the effects of active vitamin D on macrophage cholesterol deposition. METHODS AND RESULTS: We obtained macrophages from 76 obese, diabetic, hypertensive patients with vitamin D deficiency (25-hydroxyvitamin D <80 nmol/L; group A) and 4 control groups: obese, diabetic, hypertensive patients with normal vitamin D (group B; n=15); obese, nondiabetic, hypertensive patients with vitamin D deficiency (group C; n=25); and nonobese, nondiabetic, nonhypertensive patients with vitamin D deficiency (group D; n=10) or sufficiency (group E; n=10). Macrophages from the same patients in all groups were cultured in vitamin D-deficient or 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] -supplemented media and exposed to modified low-density lipoprotein cholesterol. 1,25(OH)(2)D(3) suppressed foam cell formation by reducing acetylated or oxidized low-density lipoprotein cholesterol uptake in diabetic subjects only. Conversely, deletion of the vitamin D receptor in macrophages from diabetic patients accelerated foam cell formation induced by modified LDL. 1,25(OH)(2)D(3) downregulation of c-Jun N-terminal kinase activation reduced peroxisome proliferated-activated receptor-gamma expression, suppressed CD36 expression, and prevented oxidized low-density lipoprotein-derived cholesterol uptake. In addition, 1,25(OH)(2)D(3) suppression of macrophage endoplasmic reticulum stress improved insulin signaling, downregulated SR-A1 expression, and prevented oxidized and acetylated low-density lipoprotein-derived cholesterol uptake. CONCLUSIONS: These results identify reduced vitamin D receptor signaling as a potential mechanism underlying increased foam cell formation and accelerated cardiovascular disease in diabetic subjects.

Recruitment into a long-term pediatric asthma study during emergency department visits.

OBJECTIVE: Asthma is the most common chronic illness in childhood. Recruiting children and their parents into a research study in a busy urban emergency department (ED) is challenging. The purpose of this manuscript is to describe the recruitment process and the results of our recruitment in soliciting children and their parents to participate in an ED-based asthma research study. METHODS: The data for this manuscript came from a National Heart, Lung, and Blood Institute-funded study: Study of Asthma Follow-up from the Emergency Department (SAFE). SAFE evaluated an ED-based intervention to link low-income urban children with asthma to their primary care providers. Two persons were assigned specifically to enrolling, which was done from 0700 to 2300 hours Monday through Friday. Data for the analysis come from the web-based database, the master log, and the hospital's patient database. A computerized randomization scheme chose 512 patients from all patients in the master log for more detailed demographic analyses. RESULTS: Five hundred twenty-seven subjects were enrolled between February 1999 and May 2001. There were 9188 children who presented for treatment of an acute asthma exacerbation during this interval. The number of eligible parents was similar to the predicted number. Chart reviews were conducted on a subset of patients presenting to the ED to ensure that the recruitment strategy did not bias the patients enrolled. Demographic characteristics of asthma patients were similar during enrollment and non-enrollment times. Comparison of patients who were enrolled with those who were not enrolled indicated no differences by gender, race, insurance status, age, or socioeconomic status of neighborhood residence. DISCUSSION: The high rate of enrollment was primarily due to the two dedicated enrollers. The enrollers quickly learned how to function within the ED and how to interact with both families and ED staff. Strategies identified by the enrollers as helpful in randomizing subjects included visits with the parents shortly after the physician had initiated treatment so that stability of the child had been achieved. Interacting with the child and showing concern for the comfort of both the child and parent during the ED stay were important as well. CONCLUSION: Recruiting subjects into long-term follow-up studies in the ED setting is a departure from traditional ED studies. The ED enrollment offers the distinct advantage of capturing subjects who are unlikely to present for care in other locations. We were able to successfully recruit low-income urban parents of children with asthma for study of both short-term and long-term outcomes. Careful attention to planning and then integration of enrollers into the ED setting can result in successful recruitment of patients and their parents. The enrollment process successfully captured the subjects of interest without bias.