Invasive cell phenotypes have been demonstrated in malignant transformation, but not in other diseases, such as asthma. Cellular invasiveness is thought to be mediated by transforming growth factor ...(TGF)-β1 and matrix metalloproteinases (MMPs). IL-13 is a key T(H)2 cytokine that directs many features of airway remodeling through TGF-β1 and MMPs.
We hypothesized that, in human asthma, IL-13 stimulates increased airway fibroblast invasiveness via TGF-β1 and MMPs in asthma compared with normal controls.
Fibroblasts were cultured from endobronchial biopsies in 20 subjects with mild asthma (FEV(1): 90 ± 3.6% pred) and 17 normal control subjects (FEV(1): 102 ± 2.9% pred) who underwent bronchoscopy. Airway fibroblast invasiveness was investigated using Matrigel chambers. IL-13 or IL-13 with TGF-β1 neutralizing antibody or pan-MMP inhibitor (GM6001) was added to the lower chamber as a chemoattractant. Flow cytometry and immunohistochemistry were performed in a subset of subjects to evaluate IL-13 receptor levels.
IL-13 significantly stimulated invasion in asthmatic airway fibroblasts, compared with normal control subjects. Inhibitors of both TGF-β1 and MMPs blocked IL-13-induced invasion in asthma, but had no effect in normal control subjects. At baseline, in airway tissue, IL-13 receptors were expressed in significantly higher levels in asthma, compared with normal control subjects. In airway fibroblasts, baseline IL-13Rα2 was reduced in asthma compared with normal control subjects.
IL-13 potentiates airway fibroblast invasion through a mechanism involving TGF-β1 and MMPs. IL-13 receptor subunits are differentially expressed in asthma. These effects may result in IL-13-directed airway remodeling in asthma.
Leptospirosis is a globally important, fatal disease of humans, and over 160 species of animals are associated with more than 250 bacterial serovars in 64 species, but its ecology varies regionally ...and has changed over time with expansion of human development on previously agricultural and wild land. Sporadic human cases and clusters of canine leptospirosis, primarily attributable to
serogroup Pomona, have been detected in northern California. Small mesocarnivores such as raccoons and skunks frequent peridomestic space across much of the western United States and could serve as reservoirs for human and canine leptospirosis. We aimed to summarize the prevalence of infection with pathogenic leptospires in skunk and raccoon renal and urinary samples across broad geographic zones in California, and to determine whether prevalence changed during wet and dry seasons, and as functions of host species and demographic characters. Overall, 25.6% (22/86 tested) of raccoons and 28.5% (39/137 tested) of skunks were PCR-positive for
spp. in either renal tissue or urine, with leptospiral DNA in 22.0% of kidney samples and 18.8% of urine samples from raccoons and 27.8% and 14.5% of kidney and urine samples from skunks, respectively. Raccoons from the Central California and skunks from the San Francisco Bay Area had the highest overall PCR-prevalence (35.7% and 44.4%), respectively, and adults were more likely to be PCR-positive for
spp. than juveniles. There was moderate agreement between urine and renal tissue
spp. PCR with sensitivity for both host species in renal tissue of 0.86-0.97 and 0.42-0.64 in urine. Cases of human leptospirosis are thought to be underrecognized in the continental United States and possibly increasing in some states, including California. Our data document regionally high rates of infection in common mesocarnivores, which can pose a threat to humans and dogs, revealing an important periurban epidemiological cycle.
Tumour-infiltrating CD8+ cytotoxic T cells confer favourable prognosis in colorectal cancer. The added prognostic value of other infiltrating immune cells is unclear and so we sought to investigate ...their prognostic value in two large clinical trial cohorts.
We used multiplex immunofluorescent staining of tissue microarrays to assess the densities of CD8+, CD20+, FoxP3+, and CD68+ cells in the intraepithelial and intrastromal compartments from tumour samples of patients with stage II–III colorectal cancer from the SCOT trial (ISRCTN59757862), which examined 3 months versus 6 months of adjuvant oxaliplatin-based chemotherapy, and from the QUASAR 2 trial (ISRCTN45133151), which compared adjuvant capecitabine with or without bevacizumab. Both trials included patients aged 18 years or older with an Eastern Cooperative Oncology Group performance status of 0–1. Immune marker predictors were analysed by multiple regression, and the prognostic and predictive values of markers for colorectal cancer recurrence-free interval by Cox regression were assessed using the SCOT cohort for discovery and QUASAR 2 cohort for validation.
After exclusion of cases without tissue microarrays and with technical failures, and following quality control, we included 2340 cases from the SCOT trial and 1069 from the QUASAR 2 trial in our analysis. Univariable analysis of associations with recurrence-free interval in cases from the SCOT trial showed a strong prognostic value of intraepithelial CD8 (CD8IE) as a continuous variable (hazard ratio HR for 75th vs 25th percentile 75vs25 0·73 95% CI 0·68–0·79, p=2·5 × 10−16), and of intrastromal FoxP3 (FoxP3IS; 0·71 0·64–0·78, p=1·5 × 10−13) but not as strongly in the epithelium (FoxP3IE; 0·89 0·84–0·96, p=1·5 × 10−4). Associations of other markers with recurrence-free interval were moderate. CD8IE and FoxP3IS retained independent prognostic value in bivariable and multivariable analysis, and, compared with either marker alone, a composite marker including both markers (CD8IE-FoxP3IS) was superior when assessed as a continuous variable (adjusted aHR75 vs 25 0·70 95% CI 0·63–0·78, p=5·1 × 10−11) and when categorised into low, intermediate, and high density groups using previously published cutpoints (aHR for intermediate vs high 1·68 95% CI 1·29–2·20, p=1·3 × 10−4; low vs high 2·58 1·91–3·49, p=7·9 × 10−10), with performance similar to the gold-standard Immunoscore. The prognostic value of CD8IE-FoxP3IS was confirmed in cases from the QUASAR 2 trial, both as a continuous variable (aHR75 vs 25 0·84 95% CI 0·73–0·96, p=0·012) and as a categorical variable for low versus high density (aHR 1·80 95% CI 1·17–2·75, p=0·0071) but not for intermediate versus high (1·30 0·89–1·88, p=0·17).
Combined evaluation of CD8IE and FoxP3IS could help to refine risk stratification in colorectal cancer. Investigation of FoxP3IS cells as an immunotherapy target in colorectal cancer might be merited.
Medical Research Council, National Institute for Health Research, Cancer Research UK, Swedish Cancer Society, Roche, and Promedica Foundation.
Insulin monitoring is clinically relevant for diabetes management. We present a first evaluation of an insulin immunosensor as a step towards point-of-care devices to enhance automated insulin ...delivery for T1D. The sensor is a novel, wearable, minimally invasive microneedle/microfluidic device for real-time insulin monitoring. To validate the sensor, a mid-study analysis was performed on 4 adults with T1D (43±12 years, 75% male) using insulin pumps. Insulin lispro or aspart was injected via syringe just prior to breakfast. Insulin levels were collected prior to injection and over the next 4h. Insulin quantification was performed using the participants’ serum samples extracted from venous blood. The change in real-time immunosensor-measured insulin levels was compared with values obtained by ELISA in a central lab. At each time point, a median insulin level was derived from replicate immunosensor samples. Concordance of immunosensor and ELISA insulin levels was assessed as a comparison of their changes from baseline (Figure). The mean absolute relative difference between immunosensor and venous insulin levels was 16%, 8%, and 18% at 1, 2, and 4h after the baseline measurement, respectively. The results of the 1st phase real-time insulin immunosensor evaluation are promising and may provide data to improve current insulin decay curve assumptions used in insulin-on-board approximations.
Disclosure
K.L. Wolkowicz: None. E. Vargas: None. H. Teymourian: None. F. Tehrani: None. J.E. Pinsker: Advisory Panel; Self; Medtronic. Consultant; Self; Eli Lilly and Company, Tandem Diabetes Care. Research Support; Self; Dexcom, Inc., Eli Lilly and Company, Insulet Corporation, Medtronic, Tandem Diabetes Care. Speaker’s Bureau; Self; Tandem Diabetes Care. M. Church: None. M. Piper: None. F.J. Doyle: Research Support; Self; DreaMed Diabetes, Tandem Diabetes Care, Xeris Pharmaceuticals, Inc. Stock/Shareholder; Self; Mode AGC. Other Relationship; Self; Dexcom, Inc., Insulet Corporation, Roche Diabetes Care. M. Patti: Consultant; Self; Fractyl Laboratories, Inc. Research Support; Self; Dexcom, Inc., Xeris Pharmaceuticals, Inc. Other Relationship; Self; Academy of Nutrition and Dietetics, American Diabetes Association, American Society of Metabolic and Bariatric Surgery, Endocrine Society, Insulet Corporation, King Abdullah International Medical Research Center, SUNY Downstate. L.M. Laffel: Advisory Panel; Self; Roche Diabetes Care. Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc., ConvaTec Inc., Dexcom, Inc., Insulet Corporation, Insulogic LLC, Janssen Pharmaceuticals, Inc., Lilly Diabetes, Novo Nordisk Inc., Sanofi US. J. Wang: None. E. Dassau: Consultant; Self; Eli Lilly and Company. Research Support; Self; Dexcom, Inc., DreaMed Diabetes, Tandem Diabetes Care, Xeris Pharmaceuticals, Inc. Speaker’s Bureau; Self; Roche Diabetes Care. Other Relationship; Self; Dexcom, Inc., Insulet Corporation, Roche Diabetes Care.
Funding
The Leona M. and Harry B. Helmsley Charitable Trust (2018PG-TID061); Dexcom, Inc. (IIS-2019-052)
Automated Insulin Delivery (AID) hybrid closed-loop systems have not been well studied in the context of prescribed meals. We evaluated our interoperable artificial pancreas system (iAPS) with ...scheduled meal challenges in a randomized crossover trial in an at-home setting.
Ten adults with type 1 diabetes completed two weeks of AID-based control and two weeks of conventional therapy (sensor-augmented pump/predictive low-glucose suspend) at home in random order. During each period, subjects consumed pasta or white rice as part of a complete dinner meal on six different occasions (each meal three times in random order).
The AID system increased time in range 70-180 mg/dL from 70.6 to 74.0% (3.4, 95% CI -2.3 to 9.1, p=0.22), while sensor time <70 mg/dL significantly decreased from 3.5 to 2.3% (-1.2, 95% CI -2.1 to -0.2, p=0.02). Postprandial glucose AUC difference between meals in conventional therapy was 10,919.0 mg/dL x min (95% CI 3,190.5 to 18,648.0, p=0.009) while for AID it was 6,522.1 mg/dL x min (p=0.24), highlighting the ability of AID to minimize glycemic excursions postprandially. The AID system decreased insulin delivery at 0-2h by 0.45 units (p=0.001) for pasta, while it increased at 2-4h by 0.47 units (p=0.18) for white rice (Table 1).
The AID system improved postprandial glucose control over conventional therapy in the handling of challenging meals in the at-home setting.
Disclosure
J.E. Pinsker: Advisory Panel; Self; Medtronic. Consultant; Self; Eli Lilly and Company, Tandem Diabetes Care. Research Support; Self; Dexcom, Inc., Eli Lilly and Company, Insulet Corporation, Medtronic, Tandem Diabetes Care. Speaker’s Bureau; Self; Tandem Diabetes Care. S. Deshpande: None. M. Church: None. M. Piper: None. C.C. Andre: None. J.S. Massa: None. F.J. Doyle: Research Support; Self; DreaMed Diabetes, Tandem Diabetes Care, Xeris Pharmaceuticals, Inc. Stock/Shareholder; Self; Mode AGC. Other Relationship; Self; Dexcom, Inc., Insulet Corporation, Roche Diabetes Care. D.M. Eisenberg: Consultant; Self; Barilla Center For Food and Nutrition, Italy. E. Dassau: Consultant; Self; Eli Lilly and Company. Research Support; Self; Dexcom, Inc., DreaMed Diabetes, Tandem Diabetes Care, Xeris Pharmaceuticals, Inc. Speaker’s Bureau; Self; Roche Diabetes Care. Other Relationship; Self; Dexcom, Inc., Insulet Corporation, Roche Diabetes Care.
Funding
Barilla Center for Food & Nutrition Foundation; National Institutes of Health (DP3DK104057, DP3DK113511); Harvard Accelerator; Dexcom, Inc. (IIS-2018-019)
Pregnancies in type 1 diabetes (T1D) are high-risk, and data in the U.S. are limited regarding clinical and CGM based hypoglycemia throughout pregnancy while on sensor augmented insulin pump (SAP). ...Pregnant women with T1D on insulin pump ≤ 16 wks gestation were enrolled at 3 U.S. centers and used study Dexcom G6. We analyzed episodes of severe hypoglycemia (SH) and days with > 20 hrs of CGM for biochemical hypoglycemia (BH) based on international consensus guidelines (<63 and <54 mg/dl) and frequency/duration of prolonged hypoglycemia (PH, ≥120 mins < 54 mg/dl) Twenty pregnant subjects, age 31 ± 4.5 yrs, 14 ± 8 yrs of T1D, baseline weight 77 ± 16 kg, BMI 28 ± 6.2 kg/m2, HbA1c 6.5 ± 0.7%, were studied. Gestational age at enrollment and delivery was 11.4 ± 4 and 37.7 ± 1.5 wks respectively. One subject experienced a single episode of SH with an overall incidence 7/100 patient years. Percentage of hypoglycemia and BH events per trimester are described in Table 1. BH was common in each trimester, but most frequent in the first. Eighteen events of PH occurred in four subjects (20%) lasting 120-410 mins with two events each in two, three events in one and 11 events in one subject respectively with 15 (83%) events occurring between 12am-6am. Our study shows pregnant women with T1D on SAP frequently experience level 1 hypoglycemia, and 20% experience prolonged hypoglycemia predominantly overnight.
Disclosure
R. Kaur: None. B.H. Smith: None. J.E. Pinsker: Advisory Panel; Self; Medtronic. Consultant; Self; Eli Lilly and Company, Tandem Diabetes Care. Research Support; Self; Dexcom, Inc., Eli Lilly and Company, Insulet Corporation, Medtronic, Tandem Diabetes Care. Speaker’s Bureau; Self; Tandem Diabetes Care. B. Ozaslan: None. G. O’Malley: Research Support; Self; Abbott, Dexcom, Inc. M. Trinidad: None. D. Desjardins: None. K.N. Castorino: Research Support; Self; Abbott, Dexcom, Inc., Medtronic, Mylan, Novo Nordisk Inc. C. Levister: None. C. Reid: None. S.K. McCrady-Spitzer: None. S.J. Ogyaadu: None. M. Church: None. M. Piper: None. W.K. Kremers: Research Support; Self; AstraZeneca. B. Rosenn: None. C.J. Levy: Consultant; Self; Dexcom, Inc. Employee; Spouse/Partner; Allergan plc. Research Support; Self; Abbott, Dexcom, Inc., Insulet Corporation. E. Dassau: Consultant; Self; Eli Lilly and Company. Research Support; Self; Dexcom, Inc., DreaMed Diabetes, Tandem Diabetes Care, Xeris Pharmaceuticals, Inc. Speaker’s Bureau; Self; Roche Diabetes Care. Other Relationship; Self; Dexcom, Inc., Insulet Corporation, Roche Diabetes Care. Y.C. Kudva: Research Support; Self; Dexcom, Inc., Roche Diabetes Care. Other Relationship; Self; Abbott.
Funding
National Institutes of Health (122358); Dexcom, Inc. (AP-2018-016)
The target time in range (TIR) for pregnant women with type 1 diabetes (T1D) by the International Consensus on TIR is 70% between 63-140 mg/dL. This is difficult to achieve and is rarely met in the ...literature. Insulin use increases during pregnancy, but prospective data and guidance on pump setting adjustments are limited. Insulin pump delivery and CGM data for 17 T1D women from 3 U.S. sites were prospectively collected every 2 weeks as part of the LOIS-P trial. Subjects enrolled before 17 weeks gestational age (GA) and wore personal pumps and study Dexcom G6 CGM. Changes in mean daily total, basal and bolus doses per kilogram, and TIR for every 2 weeks GA are reported, and linear mixed effects regression models are used for evaluation across trimesters. Enrollment HbA1C was 6.4±0.8%. Total daily dose increased from 0.68 to 0.73 to 0.98 U/kg during the first, second and third trimesters, respectively (p<0.01). Basal dose increased from 0.31 to 0.36 to 0.44 (p<0.01). Bolus dose increased from 0.37 to 0.37 to 0.54 (p<0.01). Daily TIR was 68%, 60% and 63% (p=0.9), with 29% of subjects achieving >70% TIR during pregnancy. Time below target was 5%, 4% and 2%. Doses trended upwards around 24 weeks GA. Postpartum doses decreased significantly. While insulin doses were increased significantly across pregnancy, most subjects did not achieve > 70% TIR. Systems that are customized to this population’s targets with changing insulin sensitivity are needed.
Disclosure
G. O’Malley: Research Support; Self; Abbott, Dexcom, Inc. B. Ozaslan: None. C. Levister: None. M. Trinidad: None. K.N. Castorino: Research Support; Self; Abbott, Dexcom, Inc., Medtronic, Mylan, Novo Nordisk Inc. D. Desjardins: None. M. Church: None. B.H. Smith: None. S.J. Ogyaadu: None. M. Piper: None. C. Reid: None. S.K. McCrady-Spitzer: None. R. Kaur: None. W.K. Kremers: Research Support; Self; AstraZeneca. F.J. Doyle: Research Support; Self; DreaMed Diabetes, Tandem Diabetes Care, Xeris Pharmaceuticals, Inc. Stock/Shareholder; Self; Mode AGC. Other Relationship; Self; Dexcom, Inc., Insulet Corporation, Roche Diabetes Care. J.E. Pinsker: Advisory Panel; Self; Medtronic. Consultant; Self; Eli Lilly and Company, Tandem Diabetes Care. Research Support; Self; Dexcom, Inc., Eli Lilly and Company, Insulet Corporation, Medtronic, Tandem Diabetes Care. Speaker’s Bureau; Self; Tandem Diabetes Care. C.J. Levy: Consultant; Self; Dexcom, Inc. Employee; Spouse/Partner; Allergan plc. Research Support; Self; Abbott, Dexcom, Inc., Insulet Corporation. B. Rosenn: None. Y.C. Kudva: Research Support; Self; Dexcom, Inc., Roche Diabetes Care. Other Relationship; Self; Abbott. E. Dassau: Consultant; Self; Eli Lilly and Company. Research Support; Self; Dexcom, Inc., DreaMed Diabetes, Tandem Diabetes Care, Xeris Pharmaceuticals, Inc. Speaker’s Bureau; Self; Roche Diabetes Care. Other Relationship; Self; Dexcom, Inc., Insulet Corporation, Roche Diabetes Care.
Funding
National Institutes of Health (R01DK120358); Dexcom, Inc. (AP-2018-016)
Pregnancies in type 1 diabetes are high risk, and data in the United States are limited regarding continuous glucose monitoring (CGM)-based hypoglycemia throughout pregnancy while on sensor-augmented ...insulin pump therapy.
Pregnant women with type 1 diabetes in the LOIS-P Study (Longitudinal Observation of Insulin use and glucose Sensor metrics in Pregnant women with type 1 diabetes using continuous glucose monitors and insulin pumps) were enrolled before 17 weeks gestation at three U.S. centers and we used their personal insulin pump and a study Dexcom G6 CGM. We analyzed data of 25 pregnant women for CGM hypoglycemia based on international consensus guidelines for percentage time <63 and 54 mg/dL, hypoglycemic events and prolonged hypoglycemia events for 24-h, daytime, and overnight periods, and severe hypoglycemia (SH) episodes.
For a 24-h period, biweekly median percentage of time <63 mg/dL ranged from 0.8% at biweek 4-5 to 3.7% at biweek 14-15 with high variability throughout pregnancy. Median percentage of time <63 and 54 mg/dL was higher overnight than daytime (
< 0.01). Hypoglycemic events occurred throughout the pregnancy, ranged 1-4 events per 2 weeks, significantly decreased after the 20th week, and occurred predominantly during daytime (
< 0.01). For overnight period, hypoglycemia and events were more concentrated from 12 to 3 am. Seven prolonged hypoglycemia events without any associated SH occurred in four participants (16%), primarily overnight. Three participants experienced a single episode of SH.
Our results suggest a higher overall risk of hypoglycemia throughout pregnancy during the overnight period with continued daytime risk of hypoglycemic events in pregnancies complicated by type 1 diabetes.
Reprogramming somatic cells into pluripotent stem cells (iPSCs) enables the study of systems in vitro. To increase the throughput of reprogramming, we present induction of pluripotency from pooled ...cells (iPPC)—an efficient, scalable, and reliable reprogramming procedure. Using our deconvolution algorithm that employs pooled sequencing of single-nucleotide polymorphisms (SNPs), we accurately estimated individual donor proportions of the pooled iPSCs. With iPPC, we concurrently reprogrammed over one hundred donor lymphoblastoid cell lines (LCLs) into iPSCs and found strong correlations of individual donors’ reprogramming ability across multiple experiments. Individual donors’ reprogramming ability remains consistent across both same-day replicates and multiple experimental runs, and the expression of certain immunoglobulin precursor genes may impact reprogramming ability. The pooled iPSCs were also able to differentiate into cerebral organoids. Our procedure enables a multiplex framework of using pooled libraries of donor iPSCs for downstream research and investigation of in vitro phenotypes.
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•Multidonor LCLs can be reprogrammed into iPSCs as a pool•The individual proportions can be easily determined from low-coverage sequencing•The individual proportions are consistent across different experimental replicates•This method contributes to the multiplexing of experimental iPSC-based models
The ability to reprogram mature cells into induced pluripotent stem cells (iPSCs) was a major discovery that enabled a plethora of downstream experimental applications. However, the reprogramming efficiency remains low, and experimental modeling of a population of many different donor iPSCs is tedious. Here, we demonstrate that reprogramming can be performed as a mixed pool of many different donor cells, and we can deconvolute donor identities and their respective proportions within the pool via low-pass DNA sequencing.
Smullen et al. report iPPC, a method for generating iPSCs from multiple donor LCLs as a pool. The individual proportions of the mixed iPSCs can be easily accessed via low-coverage next-generation sequencing. The proportions are consistent across different experimental replicates. Also, the iPSCs can be differentiated into cerebral organoids.
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