Several decades ago, a clinical condition that included severe bone overgrowth was described in a few patients in South Africa. The autosomal-recessive disease that later was named sclerosteosis was found to be caused by a mutation in the SOTS gene causing a lack of the protein sclerostin. This protein is produced by osteocytes and exerts its effect as an inhibitor of bone formation by blocking the <em>Wnt</em> signaling pathway. By the use of a monoclonal antibody that can block sclerostin a novel therapeutic pathway for rebuilding bone has been described. Preclinical studies have shown increased bone mass following subcutaneously administered anti-sclerostin antibody in animals with induced postmenopausal osteoporosis as well as in intact male rats and non-human primates. In a phase II study the efficacy and safety of an anti-sclerostin antibody, romosozumab, has been evaluated in 419 postmenopausal women for <em>12</em> months. 70, 140 or 210 mg was given subcutaneously monthly or every three months and compared to 70 mg of oral alendronate given once a week or 20 μg of teriparatide subcutaneously once daily. All dose levels of romosozumab were associated with significant increase in BMD with the most pronounced gain in the group receiving 210 mg where lumbar spine BMD increased with 11.3% from baseline. The BMD for the placebo group decreased by 0.1% while the alendronate group increased 4.1% and the teriparatide increased 7.1%. Biochemical markers revealed a transitory increase in the bone formation marker P1NP while no change in the bone resorption marker β-CTX. In comparison, teriparatide resulted in an increase for both P1NP and β-CTX for the complete study period. Even though the rapid gain in BMD is promising when considering a treatment option for osteoporosis and other conditions with bone loss, there are so far no published studies on whether anti-sclerostin can reduce the number of fractures. <em>Wnt</em> signaling might also play an important role in fracture healing with substances that causes an upregulation of the <em>Wnt</em> pathway producing enhancement of the fracture healing process. Healing of experimental fractures in various animal models have shown improvement following subcutaneously administered anti-sclerostin antibody. While there are no published reports on the potential effect of systemically administered anti-sclerostin antibodies on fracture healing in humans.

Neuronal cell fate determinants establish the identities of neurons by controlling gene expression to regulate neuronal morphology and synaptic connectivity. However, it is not understood if neuronal cell fate determinants have postmitotic functions in synapse pattern formation. Here we identify a novel role for UNC-4 homeobox protein and its corepressor UNC-37/Groucho, in tiled synaptic patterning of the cholinergic motor neurons in <i>Caenorhabditis elegans</i>. We show that <i>unc-4</i> is not required during neurogenesis but is required in the postmitotic neurons for proper synapse patterning. In contrast, <i>unc-37</i> is required in both developing and postmitotic neurons. The synaptic tiling defects of <i>unc-4</i> mutants are suppressed by <i>bar-1/β-catenin</i> mutation, which positively regulates the expression of <i>ceh-<em>12</em>/HB9</i>. Ectopic <i>ceh-<em>12</em></i> expression partly underlies the synaptic tiling defects of <i>unc-4</i> and <i>unc-37</i> mutants. Our results reveal a novel postmitotic role of neuronal cell fate determinants in synapse pattern formation through inhibiting the canonical <em>Wnt</em> signaling pathway.

Keywords: C. elegans; cell fate determinants; developmental biology; homeobox; neuroscience; pattern formation; synapse; wnt.

OBJECTIVE

To investigate the expressions of Wnt-1, beta-catenin and adenomatous polyposis coli (APC) in oral squamous cell carcinoma (OSCC).

METHODS

Surgical specimens from 66 OSCC patients were examined for Wnt-1, beta-catenin, APC and MIB-1 expressions by immunohistochemical staining.

RESULTS

Among all the 37 cases of well differentiated OSCCs, there were 30, 25 and 31 cases of high expressions of Wnt-1, APC and beta-catenin, respectively, 7, 12 and 6 cases of low expressions. Among all the 29 cases of moderate and poor differentiated OSCCs, there were 6, 9 and 11 cases of high expressions of Wnt-1, APC and beta-catenin respectively, 23, 20 and 18 cases of low expressions. Among all the 66 cases of OSCCs, there were 32 cases of high expressions of MIB-1 and 34 cases of low expressions. Expressions of Wnt-1, beta-catenin and APC showed significant difference in different differentiation of OSCC.

CONCLUSIONS

Wnt-1, beta-catenin and APC expressions were related to the differentiation of OSCC.

Background: Severe asthma is a complex disease. Transcriptomic profiling has contributed to understanding the pathogenesis of asthma, especially type-2 inflammation. However, there is still poor understanding of non-type-2 asthma, and consequently, there are limited treatment options.

Objective: The aim of this study was to identify differentially expressed genes (DEGs) and pathways in endobronchial biopsies associated with inflammatory phenotypes of severe asthma.

<strong class="sub-title"> Methods: </strong> This cross-sectional study examined endobronchial biopsies from 47 adults with severe asthma (neutrophilic asthma (NA) n = 9, eosinophilic asthma (EA) n = 22 and paucigranulocytic asthma (PGA) n = 16) and 13 healthy controls (HC). RNA was extracted and transcriptomic profiles generated (Illumina Humanref-<em>12</em> V4) and analysed using GeneSpring GX14.9.1. Pathway identification using Ingenuity Pathway Analysis.

Results: NA had the most distinct profile, with signature of 60 top-ranked DEGs (FC >±2) including genes associated with innate immunity response, neutrophil degranulation and IL-10 signalling. NA presented enrichment to pathways previously linked to neutrophilic inflammation; dendritic cell maturation, Th1, TREM1, inflammasome, Th17 and p38 MAPK, as well as novel links to neuroinflammation, NFAT and PKCθ signalling. EA presented similar transcriptomic profiles to PGA and HC. Despite the higher proportion of bacterial colonization in NA, no changes were observed in the transcriptomic profiles of severe asthma culture positive compared with severe asthma culture negative.

Conclusions & clinical relevance: NA features a distinct transcriptomic profile with seven pathways enriched in NA compared to EA, PGA and HC. All those with severe asthma had significant enrichment for SUMOylation, basal cell carcinoma signalling and Wnt/β-catenin pathways compared to HC, despite high-dose inhaled corticosteroids. These findings contribute to the understanding of mechanistic pathways in endobronchial biopsies associated with NA and identify potential novel treatment targets for severe asthma.

Keywords: Severe asthma; endobronchial biopsies; inflammatory phenotypes.

BACKGROUND

Surgery remains an integral part of the treatment of medulloblastoma. We present our experience with repeat surgery for this tumor before initiation of adjuvant therapy.

OBJECTIVE

To report what was found intraoperatively and where at time of second-look surgery and detail any postoperative events or readmissions within 90 days of surgery.

METHODS

Two separate institutional databases were queried to identify patients who underwent repeat resection of suspected residual medulloblastoma from January 2003 to January 2017.

RESULTS

We identified 51 patients (36 male, 15 female) who underwent repeat surgery. Average age at diagnosis was 8.31 years (range, 1.3-21.2). Imaging prior to repeat surgery demonstrated unequivocal residual tumor in 37 patients, but indeterminate in 14 patients. All but 1 patient had histopathologically confirmed residual tumor (50/51, 98%). The fourth ventricle was the primary site in 39 (76%) cases, compared with hemispheric in <em>12</em> cases (24%). Thirty (59%) tumors were non-<em>WNT</em>/non-SHH. All indeterminate cases (except for 1 patient) had residual tumor. Hemostatic agents were found within the resection cavity in 80% of indeterminate cases. The most common sites of residual tumor were lateral (26/39, 67%, lateral recess and/or foramen of Luschka) and roof (25/39, 64%); the superior medullary velum was the most common region of the roof (19/25, 76%). Eight (16%) patients developed new neurological deficits: cranial nerve palsies in 5 patients and posterior fossa syndrome in 3 patients.

CONCLUSIONS

Meticulous inspection of the resection cavity is necessary, paying particular attention to the roof and lateral recess. Hemostatic agents can conceal residual tumor.

Down-regulated <em>Wnt</em> signaling is involved in brain aging with declined cognitive capacity due to its modulation on neuronal function and synaptic plasticity. However, the molecular mechanisms are still unclear. In the present study, the naturally aged rat model was established by feeding rats from 6 months old to 21 months old. The cognitive capacity of aged rats was compared with young rats as the controls and the aged rats upon <em>12</em>-week exercise interventions including treadmill running, resistance exercise, and alternating exercise with resistance exercise and treadmill running. <em>Wnt</em> signaling was examined in hippocampal tissues of the rats from different groups. Results indicated that the expression of Dickkopf-1 (DKK-1) as an antagonist of <em>Wnt</em> signal pathway, the activation of GSK-3β, and the hyperphosphorylated Tau were markedly increased as the extension of age. Meanwhile, higher p-β-catenin^{Ser33, 37, Thr41} promoted neuronal degradation of aged rats. In contrast, three kinds of exercise interventions rescued the abnormal expression of DKK-1 and synaptophysin such as PSD-93 and PSD-95 in hippocampal tissues of the aged rats; especially <em>12</em>-week treadmill running suppressed DKK-1 up-regulation, GSK-3β activation, β-catenin phosphorylation, and hyperphosphorylated Tau. In addition, the down-regulated PI3K/AKT and <em>Wnt</em> signal pathways were observed in aged rats, but could be reversed by resistance exercise and treadmill running. Moreover, the increased Bax and reduced Bcl-2 levels in hippocampal tissues of aged rats were also reversed upon treadmill running intervention. Taken together, down-regulated <em>Wnt</em> signaling suppressed PI3K/Akt signal pathway, aggravated synaptotoxicity, induced neuron apoptosis, and accelerated cognitive impairment of aged rats. However, exercise interventions, especially treadmill running, can attenuate their brain aging process <i>via</i> restoring <em>Wnt</em> signaling and corresponding targets.

The transcription factor forkhead box L2 (FOXL2) regulates sex differentiation and reproductive function. Elevated levels of this transcription factor have been observed in the diseases of the uterus, such as endometriosis. However, the impact of elevated FOXL2 expression on uterine physiology remains unknown. In order to determine the consequences of altered FOXL2 in the female reproductive axis, we generated mice with over-expression of FOXL2 (FOXL2OE) by crossing Foxl2LsL/+ with the Progesterone receptor Pgrcre model. FOXL2OE uterus showed severe morphological abnormality including abnormal epithelial stratification, blunted adenogenesis, increased endometrial fibrosis, and disrupted myometrial morphology. In contrast, increasing FOXL2 levels specifically in uterine epithelium by crossing the Foxl2LsL/+ with the lactoferrin Ltficre mice resulted in the eFOXL2OE mice with uterine epithelial stratification but without defects in endometrial fibrosis and adenogenesis, demonstrating a role of the endometrial stroma in the uterine abnormalities of the FOXL2OE mice. Transcriptomic analysis of <em>12</em> weeks old Pgrcre and FOXL2OE uterus at diestrus stage showed multiple signaling pathways related with cellular matrix, <em>wnt</em>/β-catenin, and altered cell cycle. Furthermore, we found FOXL2OE mice were sterile. The infertility was caused in part by a disruption of the hypophyseal ovarian axis resulting in an anovulatory phenotype. The FOXL2OE mice failed to show decidual responses during artificial decidualization in ovariectomized mice demonstrating the uterine contribution to the infertility phenotype. These data support that aberrantly increased FOXL2 expressions in the female reproductive tract can disrupt ovarian and uterine functions.

Keywords: FOXL2; adenogenesis; fibrosis; myometrial; stratification.

Background: The present study aimed to assess the perturbation in circular RNA (circRNA)/mRNA expression profiles and a circRNA-miRNA-mRNA coexpression network involved in the potential protective effect of diosgenin (DIO) on alveolar bone loss in rats subjected to ovariectomy (OVX).

<strong class="sub-title"> Methods: </strong> The Wistar rats (female) manipulated with sham operation were classified as the SHAM group and the grouping of OVX rats administered with DIO, estradiol valerate or vehicle for <em>12</em> weeks was DIO group, EV group and OVX group respectively. Following treatments, the plasmatic levels of osteocalcin and tumor necrosis factor-alpha and the microstructure of alveolar bone were assayed. Based on microarray analyses, we identified differentially expressed (DE) circRNAs and mRNAs in alveolar bone of rats in both OVX and DIO group. The DE circRNAs and DE mRNAs involved in the bone metabolism pathway validated by RT-qPCR were considered key circRNAs/mRNAs. On the basis of these key circRNAs/mRNAs, we predicted the overlapping relative miRNAs of key circRNAs/mRNAs, and a circRNA-miRNA-mRNA network was built.

Results: DIO showed an anti-osteopenic effect on the rat alveolar bone loss induced by OVX. In total, we found 10 DE circRNAs (6 downregulated and 4 upregulated) and 614 DE mRNAs (314 downregulated and 300 upregulated) in samples of the DIO group compared with those of the OVX group. However, only one circRNA (rno_circRNA_016717) and seven mRNAs (Sfrp1, Csf1, Il1rl1, Nfatc4, Tnfrsf1a, Pik3c2g, and Wnt9b) were validated by qRT-PCR and therefore considered key circRNA/mRNAs. According to these key circRNA/mRNAs and overlapping predicted miRNAs, a coexpression network was constructed. After network analysis, one circRNA-miRNA-mRNA axis (circRNA_016717/miR-501-5p/Sfrp1) was identified.

Conclusion: The mechanism of DIO inhibiting alveolar bone loss after OVX is possibly relevant to the simultaneous inhibition of osteogenesis and osteoclastogenesis by mediating the expression of important molecules in the Wnt, PI3K, RANK/RANKL or osteoclastogenic cytokine pathways. The circRNA_016717/miR-501-5p/Sfrp1 axis may play important roles in these processes.

Keywords: Alveolar bone; CircRNAs; Coexpression network; Diosgenin; Osteoprotective effect.

Dendritic cells (DCs) are key linkages between innate immunity and acquired immunity. The antigens that promote the functions of DCs might be the effective candidates of novel vaccine. In this research, the ability of ubiquitin-conjugating enzyme (UCE), a recognized common antigens among chicken Eimeria species, to stimulate DCs of chickens were evaluated. We cloned UCE gene from Eimeria maxima (EmUCE), and its protein expression was confirmed by SDS-PAGE and western-blot. Immunofluorescence assay confirmed the binding of rEmUCE on the surface of chicken splenic-derived DCs (ChSP-DCs). Flow cytometric analysis showed that rEmUCE-treated ChSP-DCs increased MHCII, CD1.1, CD11c, CD80, and CD86 phenotypes. qRT-PCR indicated that transcript levels of maturation markers CCL5, CCR7, and CD83 in ChSP-DCs were upregulated in response to rEmUCE. Following rEmUCE treatment, chSP-DCs activated TLR signaling and inhibited <em>Wnt</em> signaling. Moreover, rEmUCE promoted DC-mediated T-cell proliferation in DC/T-cell co-incubation. Interestingly, CD3⁺/CD4⁺ T-cells were significantly enhanced when rEmUCE-treated chSP-DCs were co-incubated with T-cells. Cytokine secretion pattern of rEmUCE-stimulated ChSP-DCs revealed that the production of IL-<em>12</em> and IFN-γ was increased whereas IL-10 and TGF-β were unchanged. Likewise, the co-incubation of ChSP-DCs with T-cells indicated increased production of IFN-γ but not IL-4. Collectively, rEmUCE could polarize DCs to immunogenic phenotype and shift the immune cells towards Th1 response. Our observations provide valuable insight for future research aimed at vaccine development against avian coccidiosis.

Background: Triple-negative breast cancers (TNBCs), accounting for approximately 15% of breast cancers, lack targeted therapy. A hallmark of cancer is metabolic reprogramming, with one-carbon metabolism essential to many processes altered in tumor cells, including nucleotide biosynthesis and antioxidant defenses. We reported that folate deficiency via folic acid (FA) withdrawal in several TNBC cell lines results in heterogenous effects on cell growth, metabolic reprogramming, and mitochondrial impairment. To elucidate underlying drivers of TNBC sensitivity to folate stress, we characterized in vivo and in vitro responses to FA restriction in two TNBC models differing in metastatic potential and innate mitochondrial dysfunction.

<strong class="sub-title"> Methods: </strong> Metastatic MDA-MB-231 cells (high mitochondrial dysfunction) and nonmetastatic M-<em>Wnt</em> cells (low mitochondrial dysfunction) were orthotopically injected into mice fed diets with either 2 ppm FA (control), 0 ppm FA, or <em>12</em> ppm FA (supplementation; in MDA-MB-231 only). Tumor growth, metabolomics, and metabolic gene expression were assessed. MDA-MB-231 and M-<em>Wnt</em> cells were also grown in media with 0 or 2.2 µM FA; metabolic alterations were assessed by extracellular flux analysis, flow cytometry, and qPCR.

Results: Relative to control, dietary FA restriction decreased MDA-MB-231 tumor weight and volume, while FA supplementation minimally increased MDA-MB-231 tumor weight. Metabolic studies in vivo and in vitro using MDA-MB-231 cells showed FA restriction remodeled one-carbon metabolism, nucleotide biosynthesis, and glucose metabolism. In contrast to findings in the MDA-MB-231 model, FA restriction in the M-Wnt model, relative to control, led to accelerated tumor growth, minimal metabolic changes, and modest mitochondrial dysfunction. Increased mitochondrial dysfunction in M-Wnt cells, induced via chloramphenicol, significantly enhanced responsiveness to the cytotoxic effects of FA restriction.

Conclusions: Given the lack of targeted treatment options for TNBC, uncovering metabolic vulnerabilities that can be exploited as therapeutic targets is an important goal. Our findings suggest that a major driver of TNBC sensitivity to folate restriction is a high innate level of mitochondrial dysfunction, which can increase dependence on one-carbon metabolism. Thus, folate deprivation or antifolate therapy for TNBCs with metabolic inflexibility due to their elevated levels of mitochondrial dysfunction may represent a novel precision-medicine strategy.

Keywords: dietary folate; glycolysis; metabolomics; mitochondria; one-carbon metabolism; triple-negative breast cancer.

Aims: The aim of this prospective study was to investigate the effects of vitamin D on the expression and activity of β-catenin, as the key molecule of the Wnt/β-catenin signaling pathway, in endometriosis women.

Materials and methods: Thirty four infertile women with stage III or IV endometriosis were randomly divided to two groups. The control group received the routine treatment and the treatment group, beside the routine protocol, received 50000 IU vitamin D weekly for 12-14 weeks. Blood and endometrial tissue were collected from both groups before and after the intervention. Protein and Gene expression levels of β-catenin were assessed by Western blotting and Real-Time PCR, respectively.

Results: Compared to before intervention, the expression of active form of β-catenin reduced significantly within treatment group (p = .000), in addition, the difference between control and treatment groups (p = .012) was significant after intervention, too. Also, the ratio of active/total form of β-catenin protein expression was significantly decreased within the treatment group at the end of intervention period (p = .000).

Conclusions: It seems vitamin D can change the activity of β-catenin protein in the endometrial cells of endometriosis patients. Further studies on the therapeutic potential of vitamin D in modifying the β-catenin activity in endometriosis patients are warranted.

Clinical trial registration number: IRCT2015081823678N1.

Trial registration date: 29 September 2015.

Keywords: Endometriosis; endometrium; vitamin D; β-catenin.

Hepatocellular carcinoma (HCC) belongs to the most frequent cancer with a high death rate worldwide. Thousands of long non-coding RNAs (lncRNAs) have been confirmed to influence the development of human cancers, including HCC. Nevertheless, the biological role of PRR34 antisense RNA 1 (PRR34-AS1) in HCC remains obscure. Here, we observed via quantitative real-time reverse transcriptase polymerase chain reaction (quantitative real-time RT-PCR) that PRR34-AS1 was highly expressed in HCC cells. Functional assays revealed that PRR34-AS1 promoted HCC cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) process <i>in vitro</i> and facilitated tumor growth <i>in vivo</i>. In addition, western blot analysis and TOP Flash/FOP Flash reporter assays verified that PRR34-AS1 stimulated <em>Wnt</em>/β-catenin pathway in HCC cells. Furthermore, RNA immunoprecipitation (RIP), RNA pull-down, and luciferase reporter assays uncovered that PRR34-AS1 sequestered microRNA-296-5p (miR-296-5p) to positively modulate E2F transcription factor 2 (E2F2) and SRY-box transcription factor <em>12</em> (SOX<em>12</em>) in HCC cells. Importantly, chromatin immunoprecipitation (ChIP) and luciferase reporter assays uncovered that E2F2 transcriptionally activated PRR34-AS1 in turn. Further, rescue experiments reflected that PRR34-AS1 affected HCC progression through targeting miR-296-5p/E2F2/SOX<em>12</em>/<em>Wnt</em>/β-catenin axis. Our findings found that PRR34-AS1 elicited oncogenic functions in HCC, which indicated that PRR34-AS1 might be a novel therapeutic target for HCC.

<strong class="sub-title"> Keywords: </strong> E2F2; PRR34-AS1; SOX<em>12</em>; <em>Wnt</em>/β-catenin; hepatocellular carcinoma.

A detailed understanding of intestinal stem cell (ISC) self-renewal and differentiation is required to treat chronic intestinal diseases. However, the different models of ISC lineage hierarchy^1-6 and segregation^{7-<em>12</em>} are subject to debate. Here, we have discovered non-canonical <em>Wnt</em>/planar cell polarity (PCP)-activated ISCs that are primed towards the enteroendocrine or Paneth cell lineage. Strikingly, integration of time-resolved lineage labelling with single-cell gene expression analysis revealed that both lineages are directly recruited from ISCs via unipotent transition states, challenging the existence of formerly predicted bi- or multipotent secretory progenitors^{7-<em>12</em>}. Transitory cells that mature into Paneth cells are quiescent and express both stem cell and secretory lineage genes, indicating that these cells are the previously described Lgr5⁺ label-retaining cells⁷. Finally, <em>Wnt</em>/PCP-activated Lgr5⁺ ISCs are molecularly indistinguishable from <em>Wnt</em>/β-catenin-activated Lgr5⁺ ISCs, suggesting that lineage priming and cell-cycle exit is triggered at the post-transcriptional level by polarity cues and a switch from canonical to non-canonical <em>Wnt</em>/PCP signalling. Taken together, we redefine the mechanisms underlying ISC lineage hierarchy and identify the <em>Wnt</em>/PCP pathway as a new niche signal preceding lateral inhibition in ISC lineage priming and segregation.

With improved survivorship in medulloblastoma, there has been an increasing incidence of late complications. To date, no studies have specifically addressed the risk of radiation-associated diffuse intrinsic pontine glioma (DIPG) in medulloblastoma survivors. Query of the International DIPG Registry identified six cases of DIPG with a history of medulloblastoma treated with radiotherapy. All patients underwent central radiologic review that confirmed a diagnosis of DIPG. Six additional cases were identified in reports from recent cooperative group medulloblastoma trials (total n = <em>12</em>; ages 7 to 21 years). From these cases, molecular subgrouping of primary medulloblastomas with available tissue (n = 5) revealed only non-<em>WNT</em>, non-SHH subgroups (group 3 or 4). The estimated cumulative incidence of DIPG after post-treatment medulloblastoma ranged from 0.3-3.9%. Posterior fossa radiation exposure (including brainstem) was greater than 53.0 Gy in all cases with available details. Tumor/germline exome sequencing of three radiation-associated DIPGs revealed an H3 wild-type status and mutational signature distinct from primary DIPG with evidence of radiation-induced DNA damage. Mutations identified in the radiation-associated DIPGs had significant molecular overlap with recurrent drivers of adult glioblastoma (e.g. NRAS, EGFR, and PTEN), as opposed to epigenetic dysregulation in H3-driven primary DIPGs. Patients with radiation-associated DIPG had a significantly worse median overall survival (median 8 months; range 4-17 months) compared to patients with primary DIPG. Here, it is demonstrated that DIPG occurs as a not infrequent complication of radiation therapy in survivors of pediatric medulloblastoma and that radiation-associated DIPGs may present as a poorly-prognostic distinct molecular subgroup of H3 wild-type DIPG. Given the abysmal survival of these cases, these findings provide a compelling argument for efforts to reduce exposure of the brainstem in the treatment of medulloblastoma. Additionally, patients with radiation-associated DIPG may benefit from future therapies targeted to the molecular features of adult glioblastoma rather than primary DIPG.

Background: Scalp hair loss (alopecia) in women is a common ageing and senescing condition. It usually presents as androgenetic alopecia (AGA) or telogen effluvium (TE) and often has pronounced psychological consequences. ALRV5XR is a novel treatment aiming to regenerate a normal hair phenotype by targeting multiple molecular pathways linked to hair growth promotion and hair follicle stem cell activation. The primary objectives of this 24-week trial were to evaluate the safety and efficacy of ALRV5XR in terminal hair (TH) regrowth in women with AGA or TE.

<strong class="sub-title"> Methods: </strong> This randomised, double-blind, placebo-controlled trial was performed in a USA community clinic. Healthy women 18-65 years of age with AGA or TE of Ludwig classification I-II and Fitzpatrick skin type I-VI were enrolled. They were allocated in a 1:1 ratio into ALRV5XR or placebo treatment groups using a random number table. Masked dermatologist assessments, phototrichograms and blood samples were obtained at baseline, <em>12</em> and 24 weeks. Subjects were given a masked treatment regimen of oral capsules, shampoo, conditioner and follicle serum for daily administration. Main outcomes were absolute and per cent changes in TH density and response rates. The trial was registered with clinicaltrials.gov (<a href="http://clinicaltrials.gov/show/NCT04450602" title="See in ClinicalTrials.gov">NCT04450602</a>) and is completed.

<strong class="sub-title"> Findings: </strong> 46 subjects (23 ALRV5XR, 23 placebo) were enrolled between April 3 and October 20, 2018. Five subjects dropped out and two were non-compliant. Thirty-nine subjects completed the trial (18 ALRV5XR, 21 placebo). At 24 weeks, the absolute change in TH density improved by 30·1THs/cm² (95% CI: 15·1-45·1; p=0·0002), and the relative density increased by 19·7% (95% CI: 8·0%-31·4%; p=0·0016). The odds ratio for being a responder (≥0 change) was 2·7. Efficacy increased 133% from week <em>12</em> to 24. Efficacy outcomes were similar in AGA and TE subjects. 66·7% of the ALRV5XR group responded by regrowing 40THs/cm² or more hair. No adverse events were reported.

Interpretation: In women with AGA or TE, ALRV5XR treatment significantly increased hair regrowth without adverse events. ALRV5XR displayed a multi-fold improved efficacy and response rate when compared to published trials of standard therapy. Progressive acceleration of TH regrowth suggests regeneration of the structure and function of non-productive telogen follicles and prolonged treatment may restore a normal hair phenotype.

Keywords: Aging; Androgenetic alopecia; Botanical; COVID-19; Female pattern hair loss; Finasteride; Hair regeneration; Hair restoration; Menopause; Minoxidil; Multi-targeting; PRP; Regenerative medicine; Senescence; Stem cell; Telogen effluvium; Wnt/beta-catenin; Women's health.

Craniosynostosis (CS) is a frequent congenital anomaly featuring the premature fusion of 1 or more sutures of the cranial vault. Syndromic cases, featuring additional congenital anomalies, make up 15% of CS. While many genes underlying syndromic CS have been identified, the cause of many syndromic cases remains unknown. We performed exome sequencing of <em>12</em> syndromic CS cases and their parents, in whom previous genetic evaluations were unrevealing. Damaging de novo or transmitted loss of function (LOF) mutations were found in 8 genes that are highly intolerant to LOF mutation (<i>P</i> = 4.0 × 10^-8); additionally, a rare damaging mutation in <i>SOX11</i>, which has a lower level of intolerance, was identified. Four probands had rare damaging mutations (2 de novo) in <i>TFAP2B</i>, a transcription factor that orchestrates neural crest cell migration and differentiation; this mutation burden is highly significant (<i>P</i> = 8.2 × 10^-<em>12</em>). Three probands had rare damaging mutations in <i>GLI2</i>, <i>SOX11</i>, or <i>GPC4</i>, which function in the Hedgehog, BMP, and <em>Wnt</em> signaling pathways; other genes in these pathways have previously been implicated in syndromic CS. Similarly, damaging de novo mutations were identified in genes encoding the chromatin modifier <i>KAT6A</i>, and <i>CTNNA1</i>, encoding catenin α-1. These findings establish <i>TFAP2B</i> as a CS gene, have implications for assessing risk to subsequent children in these families, and provide evidence implicating other genes in syndromic CS. This high yield indicates the value of performing exome sequencing of syndromic CS patients when sequencing of known disease loci is unrevealing.

Background: Asthma is a chronic respiratory disease which is not curable, yet some patients experience spontaneous remission. We hypothesized that epigenetic mechanisms may be involved in asthma remission.

<strong class="sub-title"> Methods: </strong> Clinical remission (ClinR) was defined as the absence of asthma symptoms and medication for at least <em>12</em> months, and complete remission (ComR) was defined as ClinR with normal lung function and absence of airway hyperresponsiveness. We analyzed differential DNA methylation of ClinR and ComR comparing to persistent asthma (PersA) in whole blood samples (n = 72) and nasal brushing samples (n = 97) in a longitudinal cohort of well characterized asthma patients. Significant findings of whole blood DNA methylation were tested for replication in two independent cohorts, Lifelines and Epidemiological study on the Genetics and Environment of Asthma (EGEA).

<strong class="sub-title"> Results: </strong> We identified differentially methylated CpG sites associated with ClinR (7 CpG sites) and ComR (<em>12</em>9 CpG sites) in whole blood. One CpG (cg13378519, Chr1) associated with ClinR was replicated and annotated to PEX11 (Peroxisomal Biogenesis Factor 11 Beta). The whole blood DNA methylation levels of this CpG were also different between ClinR and healthy subjects. One ComR-associated CpG (cg24788483, Chr10) that annotated to TCF7L2 (Transcription Factor 7 Like 2) was replicated and associated with expression of TCF7L2 gene. One out of seven ClinR-associated CpG sites and 8 out of <em>12</em>9 ComR-associated CpG sites identified from whole blood samples showed nominal significance (P < 0.05) and the same direction of effect in nasal brushes.

Conclusion: We identified DNA methylation markers possibly associated with clinical and complete asthma remission in nasal brushes and whole blood, and two CpG sites identified from whole blood can be replicated in independent cohorts and may play a role in peroxisome proliferation and Wnt signaling pathway.

Keywords: Asthma remission; DNA methylation; Nasal brushes; Whole blood.

In this study, importance of <em>Wnt</em>-β-catenin pathway in the development of uterine cervical carcinoma was evaluated. For this purpose, the profiles (expression/methylation/deletion) of β-catenin, p-β-catenin (Y654), <em>Wnt</em>3a, and APC were studied in disease free normal cervical epithelium (n = 9), adjacent normal cervical epithelium of primary tumors (n = 70), CIN (n = 28), CACX (n = 102) samples, and two CACX cell lines (HeLa and SiHa). Immunohistochemical analysis revealed high/medium (74-95%) expression of β-catenin/p-β-catenin (Y654) and <em>Wnt</em>3a and low expression (23-26%) of APC in proliferating basal-parabasal layers contrary to differentiated spinous layer in normal cervix irrespective of HPV16 infection. The expression profile of the genes in the basal-parabasal layers did not change significantly during development of CACX. High (66%) promoter methylation of APC was seen in basal-parabasal layers and the cervical lesions (42-69%), unlike in spinous layers (25%). The promoter methylation status of APC was validated by in vitro demethylation experiments using 5-aza-dC in CACX cell lines. However, additional deletion of APC was significantly increased from CIN (<em>12</em>%) to stage I/II (40%) and became comparable in stage III/IV (48%) of the tumor. Patients with alterations (deletion/methylation) of APC and high/medium expression of <em>Wnt</em>3a/β-catenin/p-β-catenin (Y654) showed significantly poor survival. Thus our data indicate that cumulative effect of <em>Wnt</em>3a overexpression and APC inactivation are needed for overexpression of β-catenin during the development of CACX.

The molecular events driving low-grade endometrioid endometrial carcinoma (LGEC) development-like in many cancers-are incompletely understood. Hence, here we performed multiregion, comprehensive somatic molecular profiling of routinely processed formalin-fixed, paraffin-embedded (FFPE) material from 13 cases of LGEC totaling 64 minute, spatially defined cell populations ranging from presumed precursor lesions through invasive LGEC. Shared driving <i>PTEN, PIK3R1</i>, or <i>PIK3CA</i> mutations support clonal origin of the samples in each case, except for two cases with two clonally distinct neoplastic populations, consistent with unexpected multiclonality in LGEC development. Although substantial heterogeneity in driving somatic alterations was present across populations in nearly all cases, these alterations were usually clonal in a given population, supporting continued selection and clonal sweeping of driving alterations in populations with both precursor and LGEC histology. Importantly, <i>CTNNB1</i> mutational status, which has been proposed as both prognostic and predictive in LGEC, was frequently heterogeneous and subclonal, occurring both exclusively in precursor or cancer populations in different cases. Whole-transcriptome profiling of coisolated RNA from <em>12</em> lesions (from 5 cases) was robust and confirmed histologic and molecular heterogeneity, including activated <em>Wnt</em> signaling in <i>CTNNB1</i>-mutant versus wild-type populations. Taken together, we demonstrate clinically relevant multiclonality and intratumoral heterogeneity during LGEC development with important implications for diagnosis, prognosis, and therapeutic prediction. More broadly, our methodology is broadly scalable to enable high-throughput genomic and transcriptomic characterization of precursor and invasive cancer populations from routine FFPE specimens. IMPLICATIONS: Multiregion profiling of LGEC populations using a highly scalable approach demonstrates clinically relevant multiclonality and intratumoral heterogeneity.

Glioblastoma is the most common primary adult brain tumour, and despite optimal treatment, the median survival is <em>12</em>-15 months. Patients with matched recurrent glioblastomas were investigated to try to find actionable mutations. Tumours were profiled using a validated DNA-based gene panel. Copy number variations (CNVs) and single nucleotide variants (SNVs) were examined, and potentially pathogenic variants and clinically actionable mutations were identified. The results revealed that glioblastomas were <i>IDH</i>-wildtype (<i>IDH</i>^WT; <i>n</i> = 38) and <i>IDH</i>-mutant (<i>IDH</i>^MUT; <i>n</i> = 3). SNVs in <i>TSC2</i>, <i>MSH6</i>, <i>TP53</i>, <i>CREBBP</i>, and <i>IDH1</i> were variants of unknown significance (VUS) that were predicted to be pathogenic in both subtypes. <i>IDH</i>^WT tumours had SNVs that impacted RTK/Ras/PI(3)K, p53, <em>WNT</em>, SHH, NOTCH, Rb, and G-protein pathways. Many tumours had <i>BRCA1/2</i> (18%) variants, including confirmed somatic mutations in haemangioblastoma. <i>IDH</i>^WT recurrent tumours had fewer pathways impacted (RTK/Ras/PI(3)K, p53, <em>WNT</em>, and G-protein) and CNV gains (<i>BRCA2</i>, <i>GNAS</i>, and <i>EGFR</i>) and losses (<i>TERT</i> and <i>SMARCA4</i>). <i>IDH</i>^MUT tumours had SNVs that impacted RTK/Ras/PI(3)K, p53, and <em>WNT</em> pathways. VUS in <i>KLK1</i> was possibly pathogenic in <i>IDH</i>^MUT. Recurrent tumours also had fewer pathways (p53, <em>WNT</em>, and G-protein) impacted by genetic alterations. Public datasets (TCGA and GDC) confirmed the clinical significance of findings in both subtypes. Overall in this cohort, potentially actionable variation was most often identified in <i>EGFR</i>, <i>PTEN</i>, <i>BRCA1/2</i>, and <i>ATM</i>. This study underlines the need for detailed molecular profiling to identify individual GBM patients who may be eligible for novel treatment approaches. This information is also crucial for patient recruitment to clinical trials.

<AbstractText>Long non-coding RNA myocardial infarction associated transcript (MIAT) has been suspected to be associated with poor prognosis in several malignancies. This study aims to investigate the association between MIAT expression levels and prognosis of tongue squamous cell carcinoma (TSCC).</AbstractText><AbstractText>Expressions of MIAT in TSCC specimens, corresponding adjacent non-neoplastic tongue tissues, and serums collected from 116 TSCC patients were detected by Quantitative Real-Time PCR. Then, these patients were followed up for <em>12</em> months after discharge. MIAT was suppressed and upregulated in TSCC cells, and then cell invasion and epithelial-mesenchymal transition (EMT) markers were analyzed.</AbstractText><AbstractText>Myocardial infarction associated transcript expression level in TSCC specimen was upregulated compared to adjacent non-neoplastic tongue tissue and had a significant positive association with its level in serum. MIAT levels in both TSCC specimen and serum were correlated with cervical lymph node metastasis and histological grading of TSCC patients. Results of survival analysis showed that high expression levels of MIAT in both TSCC tissues and serums indicated higher mortality and shorter survival time of TSCC patients. Receiver operating characteristic curve analysis showed that there was no significant difference between MIAT in TSCC specimen and serum to differentiate death and non-death of TSCC patients. In vitro study showed that MIAT could induce invasion of TSCC cells by regulating expressions of EMT markers through activation of <em>Wnt</em>/β-catenin signaling pathway.</AbstractText><AbstractText>Upregulated MIAT promoted EMT of TSCC cells through activation of <em>Wnt</em>/β-catenin signaling pathway and indicated a poor prognosis of TSCC, which may provide us a non-invasive approach to evaluate the prognosis of TSCC.</AbstractText>

Ankylosing spondylitis (AS) is a chronic rheumatic disease that mainly affects the spinal joints (vertebrae). Spondylitis means inflammation of the spine, and ankylosing spondylitis means that bones tend to fuse. The AS causes the vertebrae to swell in the spine. Therefore, based on protein interaction network analysis, we conducted in-depth research on the molecular mechanism of key regulatory factors in the AS disease process. We carried out a differential analysis of the expression of miRNAs in disease samples and miRNAs in normal samples. Protein network interaction analysis is performed according to a group of target genes regulated by significant differentially expressed miRNAs and clustered into an interaction module. In addition, enrichment analysis of functions and pathways was performed on these modular genes. Based on the predictive analysis of multidimensional regulators, we identified a range of regulatory factors that have potential regulatory effects on AS, such as endogenous genes and transcription factors. We obtained 20 differentially expressed miRNAs and 7082 target genes and clustered into 11 modules. Enrichment results showed that these modular genes are mainly involved in the functions and pathways of protein polyubiquitination, neutrophil activation involved in immune response, and <em>Wnt</em> signaling pathway. We revealed ten transcription factors (MYC, NFKB1, and TP53). After network connectivity analysis, we obtained <em>12</em> internal drive genes (UBE2D1, CCNF, and NEDD4). These core genes are thought to be potential regulators of AS. MYC is also considered to be a core factor that inhibits SART3 phosphorylation and plays a vital role in the immunological pathogenesis of AS. The combination of the above analysis results can provide a new idea for biologists and medical scientists to study the immune pathogenesis of AS and can provide a valuable reference for subsequent treatment options.

Keywords: A key regulator; Ankylosing spondylitis; Expression of dysregulated miRNA; MYC; SART3 phosphorylation.

BACKGROUND

Human reproduction is a multifaceted process reliant on proper blastocyst implantation, placental and fetal membrane development, and delivery of a healthy baby. Multiple factors and pathways have been reported as critical machineries for cell differentiation and survival during pregnancy, and most of them involve glycogen synthase kinase (GSK) 3α/β for cell differentiation and survival and for maintaining cellular homeostasis. Several reports on GSK3's functional role exist; however, the specific role of GSK3 in reproductive tissues and its contribution to normal or abnormal parturition are still unclear. To fill this knowledge gap, a systematic review of literature was conducted to better understand the functional role of GSK3 in various intrauterine tissues during implantation, pregnancy, and parturition.

METHODS

We conducted a systematic review of English literature published between 1980 and 2017 on GSK3's expression and function reported in reproductive tissues during pregnancy using 3 electronic databases (Web of Science, Medline, and ClinicalTrials.gov). Study selection, data extraction, and quality assessment were performed in duplicate by two independent reviewers.

RESULTS

A total of 738 citations were identified; 80 were selected for full text evaluation and 25 were included for final review. GSK3's regulation and function were mostly studied in tissues and cells from placentas (<em>12</em>), fetuses (8), uteruses (6), and ovaries (2). Measurements of total GSK3 and its isoforms (α and/or β) were determined mostly by Western blot analysis. GSK3 is primarily reported as a downstream responder of protein kinase B (AKT)-, <em>Wnt</em>-, and reactive oxygen species (ROS)-related pathways where it plays a critical role in cell survival and growth in reproductive tissues.

CONCLUSIONS

GSK3 is functionally linked to blastocyst implantation, establishment of pregnancy, trophoblast migration and invasion, decidualization, and term and preterm labor. Few reports specifically studied GSK3's expression and function in any reproductive tissues; it has mostly been studied as a secondary signaler of various conserved cell signaling pathways. Lack of scientific rigor in studying GSK3's role in reproductive tissues makes this molecule's function still obscure. No studies have reported GSK3 in the cervix, and very few reports exist in myometrium and decidua. GSK3's functions are hardly studied in reproductive tissues, and several knowledge gaps have been identified that require more functional studies in reproductive biology.

Background: Although the WW-domain-containing oxidoreductase (WWOX)/Hypoxia-inducible factor 1 (HIF1) pathway is a well-known regulator of cellular glucose and energy metabolism in pathophysiological processes, its role in gestational diabetes mellitus (GDM), remains elusive. We undertook this study to determine the effect of WWOX/HIF1A signaling on the expression of glucose metabolism genes in GDM patients.

<strong class="sub-title"> Methods: </strong> Leukocytes were obtained from 135 pregnant women with (<i>n</i> = 98) or without (<i>n</i> = 37) GDM and, in turn, 3 months (<i>n</i> = 8) and 1 year (<i>n</i> = <em>12</em>) postpartum. Quantitative RT-PCR was performed to determine gene expression profiles of the WWOX/HIF1A-related genes, including those involved in glucose transport (<i>SLC2A1, SLC2A4</i>), glycolytic pathway (<i>HK2, PKM2, PFK, LDHA</i>), <em>Wnt</em> pathway (<i>DVL2, CTNNB1</i>), and inflammatory response (<i>NFKB1</i>).

Results: GDM patients displayed a significant downregulation of WWOX with simultaneous upregulation of HIF1A which resulted in approximately six times reduction in WWOX/HIF1A ratio. As a consequence, HIF1A induced genes (SLC2A1, HK2, PFK, PKM) were found to be overexpressed in GDM compared to normal pregnancy and negative correlate with WWOX/HIF1A ratio. The postpartum WWOX expression was higher than during GDM, but its level was comparable to that observed in normal pregnancy.

Conclusions: The obtained results suggest a significant contribution of the WWOX gene to glucose metabolism in patients with gestational diabetes. Decreased WWOX expression in GDM compared to normal pregnancy, and in particular reduction of WWOX/HIF1A ratio, indicate that WWOX modulates HIF1α activity in normal tissues as described in the tumor. The effect of HIF1α excessive activation is to increase the expression of genes encoding proteins directly involved in the glycolysis which may lead to pathological changes in glucose metabolism observed in gestational diabetes.

Keywords: Gestational diabetes mellitus (GDM); Glycolysis; Hypoxia-inducible factor 1α (HIF1α); WW domain-containing oxidoreductase (WWOX).