Congenital heart disease (CHD) is the most common birth defect, yet its genetic causes continue to be obscure. Fibroblast growth factor receptor 4 (FGFR4) recently emerged in a large patient exome sequencing study as a candidate disease gene for CHD and specifically heterotaxy. In heterotaxy, patterning of the left-right (LR) body axis is compromised, frequently leading to defects in the heart's LR architecture and severe CHD. FGF ligands like FGF8 and FGF4 have been previously implicated in LR development with roles ranging from formation of the laterality organ [LR organizer (LRO)] to the transfer of asymmetry from the embryonic midline to the lateral plate mesoderm (LPM). However, much less is known about which FGF receptors (FGFRs) play a role in laterality. Here, we show that the candidate heterotaxy gene FGFR4 is essential for proper organ situs in Xenopus and that frogs depleted of fgfr4 display inverted cardiac and gut looping. Fgfr4 knockdown causes mispatterning of the LRO even before cilia on its surface initiate symmetry-breaking fluid flow, indicating a role in the earliest stages of LR development. Specifically, fgfr4 acts during gastrulation to pattern the paraxial mesoderm, which gives rise to the lateral pre-somitic portion of the LRO. Upon fgfr4 knockdown, the paraxial mesoderm is mispatterned in the gastrula and LRO, and crucial genes for symmetry breakage, like coco, xnr1, and gdf3 are subsequently absent from the lateral portions of the organizer. In summary, our data indicate that FGF signaling in mesodermal LRO progenitors defines cell fates essential for subsequent LR patterning.
Macrophage migration inhibitory factor (MIF1) is a pleiotropic cytokine involved in inflammation and cancer. Genetic knockout of Mif1 in the validated N‐butyl‐N‐(4‐hydroxybutyl) nitrosamine (BBN) model of bladder cancer (BCa) resulted in stage arrest at non‐muscle‐invasive disease in prior studies. Small‐molecule inhibition of MIF1 reduced cancer‐associated outcomes, but it did not fully recapitulate genetic models. D‐dopachrome tautomerase (gene symbol DDT), commonly referred to as MIF2, is a functional homolog of MIF1, and both MIF1 and MIF2 can bind the cell surface receptor CD74 on multiple cell types to initiate a signaling cascade. It has been proposed that this interaction mediates part of the protumorigenic effects of MIF1 and MIF2 and may explain the discordance in prior studies. We hypothesized that MIF2 functions redundantly with MIF1 in BCa development and progression. The Cancer Genome Atlas (TCGA) analysis indicated MIF and DDT expression were increased in BCa patients compared to control. 4‐Iodopyridine (4‐IPP), a combined MIF1/MIF2 inhibitor, was more efficacious than ISO‐1, a MIF1‐only inhibitor, in preventing cellular proliferation in BCa cell lines. To evaluate these findings in vivo, wild‐type (WT) and Mif1−/− animals were exposed to 0.05% BBN in drinking water for 16 weeks to initiate tumorigenesis and then evaluated over the subsequent 4 weeks for tumor formation and progression in the presence or absence of 4‐IPP. 4‐IPP reduced bladder weights in WT animals and bladder weights/tumor stage in Mif1−/− animals. To determine whether MIF1/MIF2 functioned through CD74 in BCa, WT or Cd74−/− animals were used in the same BBN model. Although these animals were partially protected against BBN‐induced BCa, 4‐IPP did not enhance this effect. In conclusion, our data suggest that MIF2 mechanistically functions in a similar protumorigenic manner to MIF1, and this is at least partially through CD74. Dual inhibition of MIF homologs is more efficacious at reducing tumor burden in this model of BCa. © 2022 The Pathological Society of Great Britain and Ireland.
RhoA is critical for cytokinesis and cell-cell adhesion. While we have made significant progress in identifying the regulators of RhoA, our understanding remains incomplete. In multiple patients with heterotaxy (Htx), a disorder of left-right (LR) patterning of the internal organs, mutations in CCDC11 were identified. In Xenopus, depletion of ccdc11 causes defects in LR patterning, recapitulating the patients' phenotype. Interestingly, with ccdc11 depletion, normally monociliated cells of the Left-Right Organizer (LRO) exhibited multiple cilia per cell.While investigating this multiciliated phenotype, we unexpectedly discovered that ccdc11 is critical for cytokinesis. In addition, CCDC11 depletion alters cell-cell adhesion with loss of junctional adhesion molecules.Based on these phenotypes, reminiscent of RhoA loss of function, we demonstrate that CCDC11 colocalizes with RhoA during cytokinesis and regulates total RhoA protein levels. In conclusion, CCDC11 surprisingly connects cytokinesis and LR patterning via RhoA regulation, providing a potential mechanism for disease pathogenesis. RESULTS AND DISCUSSION Ccdc11 is important in establishing left-right asymmetryThe embryo breaks bilateral symmetry beginning at the Left-Right Organizer (LRO) [1][2][3]. Here, a subset of cells with motile monocilia drive extracellular fluid towards the left, which is sensed by immotile monocilia [4][5][6].As a consequence, dand5 (Cerl2 in mouse, coco in frog), which is initially expressed bilaterally at the margin of the LRO, is repressed on the left [2,7,8]. Repression of dand5 leads to expression of pitx2 in the left lateral plate mesoderm [1,9,10]. Pitx2 drives asymmetric organogenesis such that the heart, which initiates as a midline tube, loops to the right while other organs like the gut also undergo asymmetric morphogenesis [11].Therefore, a signaling cascade including cilia driven extracellular fluid flow, dand5, and pitx2 establish laterality of internal organs such as the heart.CCDC11 plays a key role in establishing LR asymmetry [12][13][14][15]. A number of Htx patients have CCDC11 alleles, and depletion of ccdc11 causes LR patterning defects in zebrafish and Xenopus [12][13][14][15]. Consistent with these studies, we also show that ccdc11 is important for LR patterning in Xenopus. We depleted Ccdc11 using F0 CRISPR and examined cardiac looping ( Figure 1A). We found that embryos depleted of ccdc11 showed significant cardiac looping defects (~22%) compared to uninjected controls. In addition, overexpression of human CCDC11 also showed significant cardiac looping defects (~15%) compared to uninjected controls suggesting that precise regulation of CCDC11 levels is necessary for its proper function ( Figure 1A). To place CCDC11 in the LR patterning cascade, we next examined pitx2 and coco. Using a ccdc11 morpholino oligo (MO), we found that ccdc11 knockdown led to abnormal patterns of both pitx2 (~25%) and coco (~80%) (Figures 1B-C). Therefore, using either depletion strategy, F0 CRISPR or MO, ccdc11 depletion is critica...
On univariate analysis, mpMRI predictors of ipsilateral EPE at RP were the presence of PI-RADS 5 or 4 lesion and capsular abutment/bulging (all p0.0006). Pathologic predictors (all p0.0181) included: higher GG, % overall positive cores, maximum core % involvement, maximum core % GP4, number of cores with cribriform pattern, and presence of perineural invasion on SBþTB; number of positive cores and number of positive sextants on SB. On multivariate analysis, only the presence of PI-RADS 5 lesion (OR 3.827, 95%CI 1.293-11.327, p¼0.0154) and maximum core % GP4 (OR 1.027, 95%CI 1.014-1.041, p<0.0001) on SBþTB were independently predictive.CONCLUSIONS: When selecting patients for NVB-sparing RP, balancing the achievement of safe oncologic margins with potential functional morbidity is a challenge. While the utilization of mpMRI in pre-RP staging has been generally limited due to poor sensitivity for detecting microscopic EPE, incorporation of contemporary pathologic criteria after MRI/US fusion biopsy can enhance pre-RP staging. Our data suggests that ipsilateral NVB-sparing may be considered in the absence of a PIRADS 5 lesion and with low volume GP4 involvement on MRI/US fusion biopsy.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.