Investigating the role of HOTTIP, HOXA13 and PSIP1 in cancer.

Worldwide, colorectal cancer (CRC) is the third most prevalent cancer and prostate cancer (PCa) is the second most prominent cancer in men. Treatments for both CRC and PCa have greatly improved over the last 10-20 years; however, there is still a high mortality rate for both cancers. There is therefore a great need to better understand the factors that drive disease development and progression, and therapy resistance. Long non-coding RNAs (lncRNAs) have gained attention in recent years as potential therapeutic targets for cancer. HOTTIP is one such lncRNA that has previously been implicated in CRC and PCa. HOTTIP is located on the 5’ end of the HOXA locus and regulates nearby HOXA genes including HOXA13 in cis. It has previously been shown that HOTTIP and HOXA genes are regulated by the oncogenic PSIP1 protein. Using both siRNA mediated knockdown and CRISPR-Cas9 mediated genetic knockout approaches, the role of HOTTIP, HOXA13 and PSIP1 were investigated in both CRC and PCa. Through loss of function studies, depletion of HOTTIP, HOXA13 and PSIP1 led to increased sensitivity to chemotherapeutic drugs. This has not previously been observed for HOXA13 in PCa, or HOTTIP, HOXA13 and PSIP1 in CRC. These observations suggest that further study of these genes may help with understanding therapy resistance in CRC and PCa. More in-depth characterisation of how these genes are regulated may identify methods to enhance therapy response or re-sensitise tumours to chemotherapy

Season and vitamin D status are independently associated with glucose homeostasis in pregnancy

Background: Vitamin D status and season are intrinsically linked, and both have been proposed to be associated with glucose homeostasis in pregnancy, with conflicting results. We aimed to determine if exposure to winter and low maternal 25 hydroxyvitamin D (25OHD) in early pregnancy were associated with maternal glucose metabolism.Methods: This is a secondary data analysis of 334 pregnant women enrolled in the ROLO study, Dublin. Serum 25OHD, fasting glucose, insulin and insulin resistance (HOMA-IR) were measured in early (12 weeks' gestation) and late pregnancy (28 weeks' gestation). Season of first antenatal visit was categorised as extended winter (November–April) or extended summer (May–October). Multiple linear regression models, adjusted for confounders, were used for analysis.Results: Those who attended their first antenatal visit in extended winter had lower 25OHD compared to extended summer (32.9 nmol/L vs. 44.1 nmol/L, P < 0.001). Compared to those who attended their first antenatal visit during extended summer, extended winter was associated with increased HOMA-IR in early-pregnancy (46.7%) and late pregnancy (53.7%), independent of 25OHD <30 nmol/L and confounders. Early pregnancy 25OHD <30 nmol/L and extended winter were independently associated with significantly higher fasting glucose in late pregnancy (B = 0.15 and 0.13, respectively).Conclusions: Women who attended their first antenatal visit during the months of extended winter were more likely to have raised insulin resistance in early pregnancy, which had a lasting association to 28 weeks, and was independent of 25OHD. Our novel findings imply that seasonal variation in insulin resistance may not be fully explained by differences in vitamin D status. This could reflect circannual rhythm or seasonal lifestyle behaviours, and requires further exploration.Trial registration: ISRCTN registry, ISRCTN54392969, date of registration: 22/04/2009, retrospectively registered

Evidence That a TRPA1-Mediated Murine Model of Temporomandibular Joint Pain Involves NLRP3 Inflammasome Activation

From MDPI via Jisc Publications RouterHistory: accepted 2021-10-14, pub-electronic 2021-10-23Publication status: PublishedFunder: Versus Arthritis; Grant(s): 21541This study investigates the role of transient receptor potential ankyrin 1 (TRPA1) in murine temporomandibular joint (TMJ) inflammatory hyperalgesia and the influence of the NLR family pyrin domain-containing 3 (NLRP3) inflammasome. Two distinct murine models of TMJ pain and inflammation (zymosan and CFA) were established. Spontaneous pain-like behaviours were observed as unilateral front paw cheek wipes. Ipsilateral cheek blood flow was used as a measure of ongoing inflammation, which, to our knowledge, is a novel approach to assessing real-time inflammation in the TMJ. Joint tissue and trigeminal ganglia were collected for ex vivo investigation. Both zymosan and CFA induced a time-dependent increase in hyperalgesia and inflammation biomarkers. Zymosan induced a significant effect after 4 h, correlating with a significantly increased IL-1β protein expression. CFA (50 µg) induced a more sustained response. The TRPA1 receptor antagonist A967079 significantly inhibited hyper-nociception. The NLRP3 inhibitor MCC950 similarly inhibited hyper-nociception, also attenuating inflammatory markers. In the trigeminal ganglia, CFA-induced CGRP expression showed trends of inhibition by A967079, whilst lba1 immunofluorescence was significantly inhibited by A967079 and MCC950, where the effect of TRPA1 inhibition lasted up to 14 days. Our results show that stimulation of TRPA1 is key to the TMJ pain. However, the inflammasome inhibitor exhibited similar properties in attenuating these pain-like behaviours, in addition to some inflammatory markers. This indicates that in addition to the therapeutic targeting of TRPA1, NLRP3 inhibition may provide a novel therapeutic strategy for TMJ inflammation and pain

Psip1/p52 regulates posterior Hoxa genes through activation of lncRNA Hottip

Long noncoding RNAs (lncRNAs) have been implicated in various biological functions including the regulation of gene expression, however, the functionality of lncRNAs is not clearly understood and conflicting conclusions have often been reached when comparing different methods to investigate them. Moreover, little is known about the upstream regulation of lncRNAs. Here we show that the short isoform (p52) of a transcriptional co-activator—PC4 and SF2 interacting protein (Psip1), which is known to be involved in linking transcription to RNA processing, specifically regulates the expression of the lncRNA Hottip–located at the 5’ end of the Hoxa locus. Using both knockdown and knockout approaches we show that Hottip expression is required for activation of the 5’ Hoxa genes (Hoxa13 and Hoxa10/11) and for retaining Mll1 at the 5’ end of Hoxa. Moreover, we demonstrate that artificially inducing Hottip expression is sufficient to activate the 5’ Hoxa genes and that Hottip RNA binds to the 5’ end of Hoxa. By engineering premature transcription termination, we show that it is the Hottip lncRNA molecule itself, not just Hottip transcription that is required to maintains active expression of posterior Hox genes. Our data show a direct role for a lncRNA molecule in regulating the expression of developmentally-regulated mRNA genes in cis

An examination of whether associations exist between maternal and neonatal 25OHD and infant size and adiposity at birth, 6–9 months and 2–2.5 years of age – a longitudinal observational study from the ROLO study

Background: Vitamin D status in pregnancy and offspring bone health effects are well established, yet limited knowledge exists on the effect of maternal vitamin D status on offspring size/adiposity. This study examines the association of early (13 weeks), late (28 weeks) pregnancy and neonatal (umbilical) 25-hydroxyvitamin D (25OHD) on offspring size/adiposity.Methods: This analysis included mother-infant pairs from the ROLO study at birth (n = 292), 6–9 months (n = 160) and 2–2.5 years (n = 287) postpartum.Results: Using Institute of Medicine 2011 Report criteria, 30% of women in early pregnancy and 38% in late pregnancy were at risk of vitamin D deficiency (25OHD < 30 nmol/L). Birthweight was negatively associated with early-pregnancy 25OHD (p = 0.004) and neonatal 25OHD (p < 0.001). Birth length was not associated with 25OHD. Neonatal measures of overall adiposity were negatively associated with neonatal 25OHD (p = 0.001, and p = <0.001 respectively). At 2–2.5 years there was a negative association between weight-for-age z-score and early-pregnancy 25OHD (p < 0.041).Conclusions: Maternal and neonatal 25OHD were negatively associated with offspring size/adiposity at birth and offspring weight-for-age at 2–2.5 years. Results may not reflect a general population replete in vitamin D, due to high prevalence of macrosomia and high risk of deficiency in this cohort. Improvement of pregnancy vitamin D status remains a public health concern.Trial registration: Current Controlled Trials ISRCTN54392969. 22/04/2009 retrospectively registered

Additional file 1: Table S1. of An examination of whether associations exist between maternal and neonatal 25OHD and infant size and adiposity at birth, 6–9 months and 2–2.5 years of age – a longitudinal observational study from the ROLO study

Comparison of participants at 2–2.5 years post-partum (who also had vitamin D levels from pregnancy available) and non-responders. (DOCX 15 kb

Additional file 1: Figure S1. of Season and vitamin D status are independently associated with glucose homeostasis in pregnancy

Mean early pregnancy 25OHD (nmol/L) versus month of first antenatal visit. Table S1. Maternal characteristics between included and excluded ROLO Study participant. Table S2. Dietary intakes, emotional well-being and lifestyle behaviours according to season of first antenatal visit. Table S3. Multiple Linear Regression analysis and associated percentage difference in markers of glucose metabolism (Model 2 additionally adjusted for change in 25OHD from early to late pregnancy). (DOCX 37 kb

Evidence That a TRPA1-Mediated Murine Model of Temporomandibular Joint Pain Involves NLRP3 Inflammasome Activation

This study investigates the role of transient receptor potential ankyrin 1 (TRPA1) in murine temporomandibular joint (TMJ) inflammatory hyperalgesia and the influence of the NLR family pyrin domain-containing 3 (NLRP3) inflammasome. Two distinct murine models of TMJ pain and inflammation (zymosan and CFA) were established. Spontaneous pain-like behaviours were observed as unilateral front paw cheek wipes. Ipsilateral cheek blood flow was used as a measure of ongoing inflammation, which, to our knowledge, is a novel approach to assessing real-time inflammation in the TMJ. Joint tissue and trigeminal ganglia were collected for ex vivo investigation. Both zymosan and CFA induced a time-dependent increase in hyperalgesia and inflammation biomarkers. Zymosan induced a significant effect after 4 h, correlating with a significantly increased IL-1β protein expression. CFA (50 µg) induced a more sustained response. The TRPA1 receptor antagonist A967079 significantly inhibited hyper-nociception. The NLRP3 inhibitor MCC950 similarly inhibited hyper-nociception, also attenuating inflammatory markers. In the trigeminal ganglia, CFA-induced CGRP expression showed trends of inhibition by A967079, whilst lba1 immunofluorescence was significantly inhibited by A967079 and MCC950, where the effect of TRPA1 inhibition lasted up to 14 days. Our results show that stimulation of TRPA1 is key to the TMJ pain. However, the inflammasome inhibitor exhibited similar properties in attenuating these pain-like behaviours, in addition to some inflammatory markers. This indicates that in addition to the therapeutic targeting of TRPA1, NLRP3 inhibition may provide a novel therapeutic strategy for TMJ inflammation and pain