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Current therapeutic strategies against Wilms' tumor (WT) reach 80% to 85% success rate. In spite of this, a remaining 15% to 20% of tumors relapse and are associated with increased metastasis and poor prognosis. To identify new regulators of WT progression, we screened for developmental target genes of Pax2, a key regulator of kidney development and a WT signature gene. We show that one of these target genes, calcineurin A–binding protein (CnABP), is coexpressed with Pax2 during kidney development and is overexpressed in >70% of WT samples analyzed. The CnABP gene encodes a novel protein product conserved in higher vertebrates. We show that CnABP promotes cell proliferation and migration in cell culture experiments. Biochemical analyses additionally identified an interaction between CnABP and calcineurin Aβ, the catalytic subunit of the calcium-responsive serine/threonine phosphatase calcineurin. We show that this interaction leads to the inhibition of calcineurin phosphatase activity and prevents nuclear factor of activated T-cell (NFAT) nuclear translocation. Inhibition of NFAT nuclear localization results in decreased NFAT transcriptional response. Together, these data identify a new modulator of calcineurin signaling up-regulated in WTs. (Mol Cancer Res 2009;7(6):821–31)

Objective Weight loss attempts occur as early as childhood. The impacts of weight loss attempts and weight-related stress on the occurrence of obesity during childhood remain unknown. We aimed to: (a) assess the prevalence of self-reported weight loss attempts and weight-related stress in 8-10 year-old children and (b) determine associations with adiposity 2 years later. Method Data were collected from a cohort study of 564 Canadian children aged 8-10 years, having one or both biological parents with obesity (Quebec Adipose and Lifestyle Investigation in Youth cohort). Self-reported weight loss attempts and weight-related stress were assessed at baseline in the child's questionnaire. Adiposity was measured at baseline and 2 years later using body mass index z-scores (zBMI), waist-to-height ratio (WHtR), and percentage of body fat (%BF) obtained from dual-energy x-ray absorptiometry. Linear and logistic regression analyses were used while adjusting for potential confounders. Results Forty-eight percent of children reported previous weight loss attempts and 20% reported weight-related stress. Self-reported weight loss attempts and weight-related stress were associated with higher zBMI, WHtR, and %BF 2 years later in adjusted models, although estimates were attenuated when including baseline adiposity measures. Self-reported weight loss attempts, but not weight-related stress, increased the risk of becoming overweight among children who were normal weight at baseline. Discussion Weight loss attempts are prevalent in children with parental obesity. Children reporting weight loss attempts and weight-related stress tend to have higher adiposity 2 years later and are more likely to become overweight.

Background Distinguishing lung adenocarcinoma (ADC) from squamous cell carcinoma (SCC) has a tremendous therapeutic implication. Sometimes, the commonly used immunohistochemistry (IHC) markers fail to discriminate between them, urging for the identification of new diagnostic biomarkers. Methods We performed IHC on tissue microarrays from two cohorts of lung cancer patients to analyse the expression of beta-arrestin-1, beta-arrestin-2 and clinically used diagnostic markers in ADC and SCC samples. Logistic regression models were applied for tumour subtype prediction. Parallel reaction monitoring (PRM)-based mass spectrometry was used to quantify beta-arrestin-1 in plasma from cancer patients and healthy donors. Results Beta-arrestin-1 expression was significantly higher in ADC versus SCC samples. Beta-arrestin-1 displayed high sensitivity, specificity and negative predictive value. Its usefulness in an IHC panel was also shown. Plasma beta-arrestin-1 levels were considerably higher in lung cancer patients than in healthy donors and were higher in patients who later experienced a progressive disease than in patients showing complete/partial response following EGFR inhibitor therapy. Conclusions Our data identify beta-arrestin-1 as a diagnostic marker to differentiate ADC from SCC and indicate its potential as a plasma biomarker for non-invasive diagnosis of lung cancer. Its utility to predict response to EGFR inhibitors is yet to be confirmed.

Summary During prostate development, basal and luminal cell lineages are generated through symmetric and asymmetric divisions of bipotent basal cells. However, the extent to which spindle orientation controls division symmetry or cell fate, and the upstream factors regulating this process, are still elusive. We report that GATA3 is expressed in both prostate basal progenitor and luminal cells and that loss of GATA3 leads to a mislocalization of PRKCZ, resulting in mitotic spindle randomization during progenitor cell division. Inherently proliferative intermediate progenitor cells accumulate, leading to an expansion of the luminal compartment. These defects ultimately result in a loss of tissue polarity and defective branching morphogenesis. We further show that disrupting the interaction between PRKCZ and PARD6B is sufficient to recapitulate the spindle and cell lineage phenotypes. Collectively, these results identify a critical role for GATA3 in prostate lineage specification, and further highlight the importance of regulating spindle orientation for hierarchical cell lineage organization., Graphical Abstract, Highlights • Gata3 regulates prostate lineage specification and tissue architecture • Loss of Gata3 causes aPKC mislocalization and mitotic spindle randomization • aPKC-Par6 decoupling randomizes the spindle and perturbs lineage specification • Spindle regulation prevents progenitor cell accumulation and tissue hyperplasia, In this report, Bouchard and colleagues identify a role for the transcription factor Gata3 in mitotic spindle orientation and lineage specification of prostate progenitor cells through the subcellular localization of the polarity protein aPKC. Their results underscore the importance of mitotic spindle orientation for progenitor cell homeostasis and tissue architecture.

ObjectiveTo assess whether aerobic exercise was superior to usual care in alleviating depressive symptoms in patients living with a major non-communicable disease.Data sourcesData were obtained from online databases (PubMed, PsycINFO and SPORTDiscus) as well as from reference lists. The search and collection of eligible studies was conducted up to 18 October 2018 (PROSPERO registration number CRD42017069089).Study selectionWe included interventions that compared aerobic exercise with usual care in adults who reported depressive symptoms (ie, not necessarily the clinical diagnosis of depression) and were living with a major non-communicable disease.ResultsTwenty-four studies were included in the meta-analysis (4111 patients). Aerobic exercise alleviated depressive symptoms better than did usual care (standardised mean difference (SMD)=0.50; 95% CI 0.25 to 0.76; Grading of Recommendations Assessment, Development and Evaluation: low quality). Aerobic exercise was particularly effective in alleviating depressive symptoms in cardiac patients (SMD=0.67; 95% CI 0.35 to 0.99).ConclusionAerobic exercise alleviated depressive symptoms in patients living with a major non-communicable disease, particularly in cardiac populations. Whether aerobic exercise treats clinically diagnosed depression was outside the scope of this study.

Resume La peur de l’accouchement est tres frequente chez les femmes enceintes d’un premier enfant. Elle semble liee a des consequences specifiques a l’accouchement et en post-partum, dont l’apparition de symptomes post-traumatiques (SPT), de meme que le blues ou la depression post-partum (DPP). Or, la plupart des etudes realisees reposent sur des echantillons tres restreints, montrent des resultats contradictoires ou tiennent peu compte des interactions entre les facteurs impliques. Objectif Cette etude vise a verifier si la peur de l’accouchement de femmes enceintes nullipares est liee au deroulement de l’accouchement et a la presence de consequences post-partum (c’est-a-dire symptomes de DPP et de SPT). Methode Au total, 176 femmes ont repondu a deux temps de mesure (pre- et post-partum). Resultats La peur de l’accouchement est liee a la douleur percue a la naissance chez les femmes accouchant par voie vaginale, en l’absence d’anesthesie. Elle est aussi liee aux symptomes de DPP et de SPT, independamment du recours ou non a l’anesthesie. La peur apparait aussi liee fortement aux symptomes de SPT chez les femmes ayant vecu une cesarienne non planifiee. Les symptomes de SPT post-partum pourraient jouer un role mediateur dans le lien unissant la peur d’accoucher et la depression post-partum. Conclusions Ces resultats supportent la pertinence de tenir compte de la peur de l’accouchement et de la douleur percue en lien avec les symptomes de SPT et de DPP aupres de nullipares. La peur de l’accouchement pourrait rendre l’experience de la naissance plus negative et predisposer au SPT et a la DPP post-partum. D’autres etudes sont necessaires pour preciser ces relations.

The cognitive approach of obsessive-compulsive disorder (OCD) has suggested a bidirectional link between emotions and cognitions. Few studies have looked at a link between those two components. Although some studies tend to show a relationship between emotions and OCD, no study has looked into the relationship between emotions, cognitions, and behaviors over the course of a cognitive therapy. The present case series examines the relationship between cognitive, behavioral, and emotional processes over the course of an inference-based therapy (IBT) in OCD clients. More precisely, we looked at how emotions and OCD symptoms influence each other and how they influence each other over time (through therapy). Clients completed daily diaries rating key emotions, behaviors, and beliefs over the course of treatment. A longitudinal analysis based on an event-based approach was used to maximize the potential of longitudinal data. Results showed that anxiety, sadness, and joy share similar trajectories with beliefs an...

Objectives Anxiety has been shown to be associated with poor outcomes in people with diabetes. However, there has been little research which has specifically examined whether diabetes mellitus is associated with an increased likelihood of co-morbid anxiety. The aim of this systematic review and meta-analysis was to determine whether people with diabetes are more likely to have anxiety disorders or elevated anxiety symptoms than people who do not have diabetes. Methods A systematic review was performed by three independent reviewers who searched for articles that examined the association between anxiety and diabetes in adults 16 or older. Those studies that met eligibility criteria were put forward for meta-analysis using a random-effects model. Results A total of twelve studies with data for 12,626 people with diabetes were eligible for inclusion in the systematic review and meta-analysis. Significant and positive associations were found for diabetes with both anxiety disorders, 1.20 (1.10–1.31), and elevated anxiety symptoms, 1.48 (1.02–1.93). The pooled OR for all studies that assessed anxiety was 1.25 (1.10–1.39). Conclusions Results from this meta-analysis provide support that diabetes is associated with an increased likelihood of having anxiety disorders and elevated anxiety symptoms.

Aims There is an increasing interest in single-item self-rated indicators of perceived health and control status in people with chronic illnesses such as diabetes. However, self-rated measures can be associated with indicators of psychological status. The aim of this paper is to explore the association of anxiety, depression, and diabetes distress with self-rated diabetes control. Methods Telephone interviews were conducted with 1,787 people with type 2 diabetes taking oral hypoglycemic medication. Diabetes control, health behaviors, and outcomes, anxiety, depression, and diabetes distress were assessed by standardized questionnaires. Self-reported diabetes control was modeled using logistic regression. Results The best fit logistic regression model for self-rated poor diabetes control was a model that incorporated diabetes distress. When adjusted for age, sex, and all other health behaviors and outcomes, poor diabetes control was most associated with diabetes distress, physical inactivity, being overweight, and poor eating habits. Conclusions Results from this study indicate that poor self-rated diabetes control shares the strongest associations with diabetes-specific distress along with perceptions of diabetes-specific healthcare behaviors and outcomes.

Lower maternal plasma volume expansion was found in idiopathic intrauterine growth restriction (IUGR) but the link remains to be elucidated. An animal model of IUGR was developed by giving a low-sodium diet to rats over the last week of gestation. This treatment prevents full expansion of maternal circulating volume and the increase in uterine artery diameter, leading to reduced placental weight compared to normal gestation. We aimed to verify whether this is associated with reduced remodeling of uteroplacental circulation and placental hypoxia. Dams were divided into two groups: IUGR group and normal-fed controls. Blood velocity waveforms in the main uterine artery were obtained by Doppler sonography on days 14, 18 and 21 of pregnancy. On day 22 (term = 23 days), rats were sacrificed and placentas and uterine radial arteries were collected. Diameter and myogenic response of uterine arteries supplying placentas were determined while expression of hypoxia-modulated genes (HIF-1α, VEGFA and VEGFR2), apoptotic enzyme (Caspase -3 and -9) and glycogen cells clusters were measured in control and IUGR term-placentas. In the IUGR group, impaired blood velocity in the main uterine artery along with increased resistance index was observed without alteration in umbilical artery blood velocity. Radial uterine artery diameter was reduced while myogenic response was increased. IUGR placentas displayed increased expression of hypoxia markers without change in the caspases and increased glycogen cells in the junctional zone. The present data suggest that reduced placental and fetal growth in our IUGR model may be mediated, in part, through reduced maternal uteroplacental blood flow and increased placental hypoxia.

Cdx and Hox gene families encode homeodomain-containing transcription factors involved in anterior-posterior vertebral patterning. Although Cdx proteins are direct transcriptional regulators of Hox gene expression, both Hox and Cdx proteins are known to interact with other homeodomain transcription factors, leading us to speculate that Cdx and Hox proteins may also interact physically. In testing this, we found that that Cdx1 is indeed capable of associating with a subset of Hox proteins. This interaction is localized to the homeodomain region of both classes of proteins, is reliant on specific arginine residues in helix I of the Hox homeodomain, and is further modulated by N-terminal Hox sequences. More promiscuous interactions were seen with Hox proteins expressed in vivo, suggestive of bridging factors or post-translational modifications. Finally, we demonstrate that this interaction modulates Cdx-Hox transcriptional activity on a Hox-responsive element. This study is the first example of Cdx-Hox protein interactions and suggests that such complexes may modulate Hox and/or Cdx function.

Resume La peur de l’accouchement, caracteristique centrale de la tocophobie, est frequente chez les femmes enceintes de leur premier enfant. Elle est associee a des consequences majeures comme la depression pre- et post-natale. Malgre cela, peu d’instruments valides sont disponibles, notamment en francais, pour evaluer ce construit. Cet article comporte les resultats de deux etudes ayant pour but d’adapter et de valider une echelle mesurant l’intensite de la peur de l’accouchement chez des femmes enceintes francophones, soit le Traumatic Event Scale (TES) adapte a la peur de l’accouchement (Soderquist et al., 2004 [21] ). La premiere presente une etape preliminaire visant a evaluer la qualite de cet instrument, de meme que sa fidelite et sa validite (convergente et factorielle) aupres d’un echantillon de meres ayant un enfant âge de 36 mois et moins (n = 65). La seconde etude verifie les proprietes psychometriques d’une version ajustee de l’instrument aupres d’un echantillon de femmes enceintes primipares (n = 204). Les resultats de l’etude 1 revelent des proprietes psychometriques adequates et une solution factorielle en cinq facteurs. Les resultats de l’etude 2 reproduisent cette solution sur la base d’analyses factorielles confirmatoires. Ces resultats confirment les qualites psychometriques de cet instrument. Une discussion entourant les contextes et avantages d’utilisation de cet outil est presentee.

Aims Diabetes control is a multifaceted process involving successful adherence to a self-care regimen as indicated by improved health outcomes. The aim of this study was to ascertain the construct validity of self-reported diabetes control in a population-based survey. Methods This study assessed 1848 participants with type 2 diabetes who took part in the Montreal Diabetes Health and Wellbeing Study in Quebec, Canada. Participants were administered the diabetes complications index as well as sociodemographic and health questions. Results Fair/poor diabetes control was associated with being less likely to check blood glucose weekly, being less likely to drink alcohol, being more likely to report being physically inactive, reporting fair/poor eating habits, being obese and having 1 or more diabetes complications. When all variables were included in a regression model the two variables most strongly associated with poor fair/poor diabetes control were reporting fair/poor eating habits (odds ratio 1.36, 95% CI 1.00–1.85) and having 2 or more diabetes complications (odds ratio 1.60, 95% CI 1.06–2.40). Conclusions Results from this study indicate that self-rated diabetes control has associations with diabetes-specific self-care behaviours and outcomes, and is a general indicator of self-care and diabetes-related complications in a population-based survey.

It is well established that Hox genes are key players in specifying positional identity along the anterior-posterior axis and are targets of diverse transcription factors implicated in axial patterning. Members of the CDX family (CDX1, -2, and -4) are among such effectors of Hox expression as pertains to vertebral patterning in the mouse. Cdx members are themselves targets of signaling molecules that are also implicated in axial patterning, including retinoic acid (RA) and certain members of Wnt and fibroblast growth factor families. In this regard, we have previously shown that, in the mouse, Cdx1 is directly regulated by RA at the late primitive streak stage (embryonic day (E) 7.5) through a RA response element in the proximal Cdx1 promoter. At E8.5, Cdx1 expression remains essentially limited to the posterior embryo. RA, however, is excluded from the caudal embryo at this later stage, but is found in a more anterior domain encompassing the prospective trunk region. These observations suggest the existence of a repressor mechanism that prevents expression of Cdx1 in these anterior domains of retinoid signaling at E8.5. In the present study, we present evidence suggesting that chicken ovalbumin upstream promoter-transcription factor (COUP-TF) members antagonize RA-induced Cdx1 expression by competing with retinoid X receptor-retinoic acid receptor heterodimers for binding to the Cdx1 RA response element. Consistent with this, in situ hybridization analysis revealed that COUP-TFs are highly expressed in the anterior embryo in domains where Cdx1 transcripts are excluded. Together with other data, these findings suggest a model by which COUP-TF expression is induced by RA in the trunk region as a negative feedback mechanism to restrict Cdx1 expression to the caudal embryo.

Neural crest cells (NCC) are a transient migratory cell population that generates diverse cell types such as neurons and glia of the enteric nervous system (ENS). Via an insertional mutation screen for loci affecting NCC development in mice, we identified one line—named TashT—that displays a partially penetrant aganglionic megacolon phenotype in a strong male-biased manner. Interestingly, this phenotype is highly reminiscent of human Hirschsprung’s disease, a neurocristopathy with a still unexplained male sex bias. In contrast to the megacolon phenotype, colonic aganglionosis is almost fully penetrant in homozygous TashT animals. The sex bias in megacolon expressivity can be explained by the fact that the male ENS ends, on average, around a “tipping point” of minimal colonic ganglionosis while the female ENS ends, on average, just beyond it. Detailed analysis of embryonic intestines revealed that aganglionosis in homozygous TashT animals is due to slower migration of enteric NCC. The TashT insertional mutation is localized in a gene desert containing multiple highly conserved elements that exhibit repressive activity in reporter assays. RNAseq analyses and 3C assays revealed that the TashT insertion results, at least in part, in NCC-specific relief of repression of the uncharacterized gene Fam162b; an outcome independently confirmed via transient transgenesis. The transcriptional signature of enteric NCC from homozygous TashT embryos is also characterized by the deregulation of genes encoding members of the most important signaling pathways for ENS formation—Gdnf/Ret and Edn3/Ednrb—and, intriguingly, the downregulation of specific subsets of X-linked genes. In conclusion, this study not only allowed the identification of Fam162b coding and regulatory sequences as novel candidate loci for Hirschsprung’s disease but also provides important new insights into its male sex bias., Author Summary Hirschsprung’s disease (also known as aganglionic megacolon) is a severe congenital defect of the enteric nervous system (ENS) resulting in complete failure to pass stools. It is characterized by the absence of neural ganglia (aganglionosis) in the distal gut due to incomplete colonization of the embryonic intestines by neural crest cells (NCC), the ENS precursors. Hirschsprung’s disease has an incidence of 1 in 5000 newborns and a 4:1 male sex bias. Although many genes have been associated with this complex genetic disease, most of its heritability as well as its male sex bias remain unexplained. Here, we describe an insertional mutant mouse line (“TashT”) in which virtually all homozygotes display colonic aganglionosis due to defective migration of enteric NCC, but in which only a subset of homozygotes develops megacolon. Surprisingly, this group is almost exclusively male. The TashT ENS defect stems, at least in part, from the disruption of long-range interactions between evolutionarily conserved elements with silencer activity and Fam162b, resulting in NCC-specific upregulation of this uncharacterized protein coding gene. Global analysis of gene expression further revealed that several hundreds of genes are significantly deregulated in TashT enteric NCC. Interestingly, this dataset includes multiple X-linked candidate genes potentially underlying the male sex bias. Taken together, our data pave the way for a clearer understanding of the intriguing male sex bias of Hirschsprung’s disease.

Current therapeutic strategies against Wilms' tumor (WT) reach 80% to 85% success rate. In spite of this, a remaining 15% to 20% of tumors relapse and are associated with increased metastasis and poor prognosis. To identify new regulators of WT progression, we screened for developmental target genes of Pax2, a key regulator of kidney development and a WT signature gene. We show that one of these target genes, calcineurin A–binding protein (CnABP), is coexpressed with Pax2 during kidney development and is overexpressed in >70% of WT samples analyzed. The CnABP gene encodes a novel protein product conserved in higher vertebrates. We show that CnABP promotes cell proliferation and migration in cell culture experiments. Biochemical analyses additionally identified an interaction between CnABP and calcineurin Aβ, the catalytic subunit of the calcium-responsive serine/threonine phosphatase calcineurin. We show that this interaction leads to the inhibition of calcineurin phosphatase activity and prevents nuclear factor of activated T-cell (NFAT) nuclear translocation. Inhibition of NFAT nuclear localization results in decreased NFAT transcriptional response. Together, these data identify a new modulator of calcineurin signaling up-regulated in WTs. (Mol Cancer Res 2009;7(6):821–31)

The Cdx transcription factors regulate anterior–posterior (AP) vertebral patterning, at least in part, through direct regulation of Hox gene expression. Analysis of allelic series of Cdx mutant mice suggests functional overlap between these family members. However, the lack of a Cdx2 null mutant makes these analyses incomplete. Moreover, Hox proteins are sometimes redundant, making it difficult to discern whether Cdx members regulate identical Hox target genes in a redundant manner, or whether they regulate separate Hox genes which then converge on events related to vertebral patterning. To more directly assess this question, we developed a “knock in” model whereby Cdx2 was substituted for Cdx1. Consistent with functional redundancy Cdx2 “knock-in” mice exhibited perfect complementation of the Cdx1-null phenotype, as evidenced by the lack of skeletal defects or altered expression of Hox genes typically impacted by Cdx1 loss-of-function. It has been proposed that vertebral AP patterning is reliant on a gradient of the sum total of Cdx proteins, a posit that is consistent with functional redundancy between Cdx family members. To further assess this, we generated a gain-of-function model using BAC trangenesis to alter Cdx1 dosage. Cdx1 BAC transgenic mice overexpressed Cdx1 mRNA and protein, and fully complemented the Cdx1 null allele. However, gain of Cdx1 dosage via this BAC transgene in an otherwise wild type background had no discernible effects on vertebral patterning or Hox gene expression, suggesting that a moderate alteration in the Cdx protein gradient is of no consequence.

The rising incidence of cancers affecting the kidney emphasizes the need to identify the molecular pathways involved in the initiation and progression of kidney tumors in order to counter this phenomenon. For many years, genes belonging to the PAX family have been the focus of intensive studies in the fields of organogenesis and tumorigenesis. PAX2 and PAX8 encode transcription factors essential for embryonic kidney development. Transcriptionnal repression of these factors is, however, required to allow terminal differentiation of renal epithelia. In human, maintenance and reactivation of PAX2/8 expression are frequently observed in cases of Wilm's tumor and renal cell carcinoma. The precise role of PAX2/8 in kidney cancer is still elusive but results from several studies suggest the exertion of common functions during organogenesis and tumorigenesis of the kidney. Moreover, many members of the PAX family are involved in similar cellular processes such as differentiation/proliferation, motility and apoptosis. Thus, by comparing the functions exerted by PAX factors in several types of cancers should be useful to better define the specific contribution of PAX2/8 to kidney tumorigenesis.

The Cdx1 gene product is essential for normal anterior-posterior vertebral patterning. Expression of Cdx1 is regulated by several pathways implicated in anterior-posterior patterning events, including retinoid and Wnt signaling. We have previously shown that retinoic acid plays a key role in early stages of Cdx1 expression at embryonic day 7.5 (E7.5), while both Wnt3a signaling and an autoregulatory loop, dependent on Cdx1 itself, are involved in later stages of expression (E8.5 to E9.5). This autoregulation is reflected by the ability of Cdx1 to affect expression from proximal Cdx1 promoter sequences in tissue culture. However, this region is devoid of a demonstrable Cdx response element(s). We have now found that Cdx1 and LEF1, a nuclear effector of Wnt signaling, synergize to induce expression from the Cdx1 promoter through previously documented LEF/T-cell factor response elements. We also found a direct physical interaction between the homeodomain of Cdx1 and the B box of LEF1, suggesting a basis for this synergy. Consistent with these observations, analysis of Cdx1 Wnt3a vt compound mutants demonstrated that Wnt and Cdx1 converged on Cdx1 expression and vertebral patterning in vivo. Further data suggest that Cdx-high-mobility group box interactions might be involved in a number of additional pathways. Somites are derived from segmentation of the paraxial mesoderm in the caudal embryo and subsequently differentiate into the dermamyotome and sclerotome, the latter being the anlage of the vertebrae. Many vertebrae exhibit morphological differences along the anterior-posterior axis, such as the ribs characteristic of thoracic vertebrae. These distinct morphological characteristics are indicative of patterning events which dictate vertebral identity along the anterior-posterior axis. A number of signaling molecules, such as retinoic acid, are well documented to affect vertebral anterior-posterior patterning. All such effectors typically impact on the expression of Hox genes, and a wealth of gain- and loss-of-function experiments clearly demonstrate a critical role for Hox gene products in vertebral patterning (see, e.g., references 9, 11, 13, 19, and 27). The 39 murine Hox genes are distributed in four clusters, Hoxa to Hoxd, which have likely evolved by duplication of an ancestral complex related to the HOM-C genes of Drosophila melanogaster (17, 18, 20). In the mouse, Hox expression is initiated at embryonic day 7.5 (E7.5) in the primitive streak, with transcripts subsequently expanding anteriorly in the neural tube and mesoderm to eventually reach a predetermined rostral limit (14, 45, 50). The onset and rostral limit of expression are generally related to the location of a given Hox gene within its complex, with more 3 members initiated earlier and reaching more rostral limits of expression than 5 paralogs. This results in staggered domains of Hox expression along the anterior-posterior axis, which have been suggested to comprise a Hox code (5, 23, 33).

Wilms' tumor (WT) is a classical cancer type that arises from abnormal differentiation of kidney progenitor cells, and occurs at a frequency of 1 in 10,000 live births, accounting for approx. 90% of childhood kidney cancer. Abnormally high levels of Pax2 (paired-box protein 2), a key regulator of kidney development, have been observed in both WT and renal cell carcinoma, which correlate with proliferation and increased invasiveness. Therefore the misexpression of Pax2 and its target genes may play an important role in tumor initiation and/or progression. To test this, we screened for target genes of Pax2 by cDNA microarray in the embryonic kidney. We identified CnABP (Calcineurin A Binding Protein), a novel gene under Pax2 regulation. In situ hybridization indicates that CnABP coexpresses with Pax2 in the condensing mesenchyme, the abnormal differentiation of which gives rise to WT. Furthermore, expression analysis by quantitative PCR indicates that CnABP is overexpressed in more than 70% of Wilms' tumors. Interestingly, in the proportion of tumors with upregulated PAX2 expression, more than 80% also overexpress CnABP. We characterized CnABP as a membrane-anchored protein that primarily promotes cell proliferation and migration. Mediators of these activities were investigated by Yeast-two-hybrid and immunoprecipitation, which identified an interaction between CnABP and Calcineurin A, the catalytic subunit of a calcium-responsive serine/threonine phosphatase Calcineurin. We showed that CnABP modulates phosphatase activities of Calcineurin, which consequently inhibits calcium/Calcineurin-dependent NFAT nuclear translocation. We further demonstrated that the inhibition of NFAT nuclear localization results in reduced NFAT-specific transcriptional activity. Components of the Calcineurin complex have been reported as differentially expressed genes distinguishing recurrent from non-recurrent Wilms' tumors. This is in line with the evidence we presented, as CnABP is upregulated in Wilms' tumors and is shown to promote proliferation and migration. Citation Information: Cancer Res 2009;69(23 Suppl):A54.