Systematic transcriptome profiling of hPSC-derived osteoblasts unveils CORIN's mastery in governing osteogenesis through CEBPD modulation.

The commitment of stem cells to differentiate into osteoblasts is a highly regulated and complex process that involves the coordination of extrinsic signals and intrinsic transcriptional machinery. While rodent osteoblastic differentiation has been extensively studied, research on human osteogenesis has been limited by cell sources and existing models. Here, we systematically dissect human pluripotent stem cell-derived osteoblasts to identify functional membrane proteins and their downstream transcriptional networks involved in human osteogenesis. Our results reveal an enrichment of type II transmembrane serine protease CORIN in humans but not rodent osteoblasts. Functional analyses demonstrated that CORIN depletion significantly impairs osteogenesis. Genome-wide chromatin immunoprecipitation enrichment and mechanistic studies show that p38 MAPK-mediated CCAAT enhancer binding protein delta (CEBPD) upregulation is required for CORIN-modulated osteogenesis. Contrastingly, the type I transmembrane heparan sulfate proteoglycan SDC1 enriched in mesenchymal stem cells exerts a negative regulatory effect on osteogenesis through a similar mechanism. Chromatin immunoprecipitation-seq, bulk and single-cell transcriptomes, and functional validations indicated that CEBPD plays a critical role in controlling osteogenesis. In summary, our findings uncover previously unrecognized CORIN-mediated CEBPD transcriptomic networks in driving human osteoblast lineage commitment.

In vivo evidence for GDP-fucose transport in the absence of transporter SLC35C1 and putative transporter SLC35C2.

Slc35c1 encodes an antiporter that transports GDP-fucose into the Golgi and returns GMP to the cytoplasm. The closely related gene Slc35c2 encodes a putative GDP-fucose transporter and promotes Notch fucosylation and Notch signaling in cultured cells. Here, we show that HEK293T cells lacking SLC35C1 transferred reduced amounts of O-fucose to secreted epidermal growth factor-like repeats from NOTCH1 or secreted thrombospondin type I repeats from thrombospondin 1. However, cells lacking SLC35C2 did not exhibit reduced fucosylation of these epidermal growth factor-like repeats or thrombospondin type I repeats. To investigate SLC35C2 functions in vivo, WW6 embryonic stem cells were targeted for Slc35c2. Slc35c2[-/-] mice were viable and fertile and exhibited no evidence of defective Notch signaling during skeletal or T cell development. By contrast, mice with inactivated Slc35c1 exhibited perinatal lethality and marked skeletal defects in late embryogenesis, typical of defective Notch signaling. Compound Slc35c1[-/-]Slc35c2[-/-] mutants were indistinguishable in skeletal phenotype from Slc35c1[-/-] embryos and neonates. Double mutants did not exhibit the exacerbated skeletal defects predicted if SLC35C2 was functionally important for Notch signaling in vivo. In addition, NOTCH1 immunoprecipitated from Slc35c1[-/-]Slc35c2[-/-] neonatal lung carried fucose detected by binding of Aleuria aurantia lectin. Given that the absence of both SLC35C1, a known GDP-fucose transporter, and SLC35C2, a putative GDP-fucose transporter, did not lead to afucosylated NOTCH1 nor to the severe Notch signaling defects and embryonic lethality expected if all GDP-fucose transport were abrogated, at least one more mechanism of GDP-fucose transport into the secretory pathway must exist in mammals.

Patient-derived iPSCs link elevated mitochondrial respiratory complex I function to osteosarcoma in Rothmund-Thomson syndrome.

Rothmund-Thomson syndrome (RTS) is an autosomal recessive genetic disorder characterized by poikiloderma, small stature, skeletal anomalies, sparse brows/lashes, cataracts, and predisposition to cancer. Type 2 RTS patients with biallelic RECQL4 pathogenic variants have multiple skeletal anomalies and a significantly increased incidence of osteosarcoma. Here, we generated RTS patient-derived induced pluripotent stem cells (iPSCs) to dissect the pathological signaling leading to RTS patient-associated osteosarcoma. RTS iPSC-derived osteoblasts showed defective osteogenic differentiation and gain of in vitro tumorigenic ability. Transcriptome analysis of RTS osteoblasts validated decreased bone morphogenesis while revealing aberrantly upregulated mitochondrial respiratory complex I gene expression. RTS osteoblast metabolic assays demonstrated elevated mitochondrial respiratory complex I function, increased oxidative phosphorylation (OXPHOS), and increased ATP production. Inhibition of mitochondrial respiratory complex I activity by IACS-010759 selectively suppressed cellular respiration and cell proliferation of RTS osteoblasts. Furthermore, systems analysis of IACS-010759-induced changes in RTS osteoblasts revealed that chemical inhibition of mitochondrial respiratory complex I impaired cell proliferation, induced senescence, and decreased MAPK signaling and cell cycle associated genes, but increased H19 and ribosomal protein genes. In summary, our study suggests that mitochondrial respiratory complex I is a potential therapeutic target for RTS-associated osteosarcoma and provides future insights for clinical treatment strategies.

Biallelic variants in CRIPT cause a Rothmund-Thomson-like syndrome with increased cellular senescence.

PURPOSE: Rothmund-Thomson syndrome (RTS) is characterized by poikiloderma, sparse hair, small stature, skeletal defects, cancer, and cataracts, resembling features of premature aging. RECQL4 and ANAPC1 are the 2 known disease genes associated with RTS in >70% of cases. We describe RTS-like features in 5 individuals with biallelic variants in CRIPT (OMIM 615789). METHODS: Two newly identified and 4 published individuals with CRIPT variants were systematically compared with those with RTS using clinical data, computational analysis of photographs, histologic analysis of skin, and cellular studies on fibroblasts. RESULTS: All CRIPT individuals fulfilled the diagnostic criteria for RTS and additionally had neurodevelopmental delay and seizures. Using computational gestalt analysis, CRIPT individuals showed greatest facial similarity with individuals with RTS. Skin biopsies revealed a high expression of senescence markers (p53/p16/p21) and the senescence-associated ß-galactosidase activity was elevated in CRIPT-deficient fibroblasts. RECQL4- and CRIPT-deficient fibroblasts showed an unremarkable mitotic progression and unremarkable number of mitotic errors and no or only mild sensitivity to genotoxic stress by ionizing radiation, mitomycin C, hydroxyurea, etoposide, and potassium bromate. CONCLUSION: CRIPT causes an RTS-like syndrome associated with neurodevelopmental delay and epilepsy. At the cellular level, RECQL4- and CRIPT-deficient cells display increased senescence, suggesting shared molecular mechanisms leading to the clinical phenotypes.

Mutations in ANAPC1, Encoding a Scaffold Subunit of the Anaphase-Promoting Complex, Cause Rothmund-Thomson Syndrome Type 1.

Rothmund-Thomson syndrome (RTS) is an autosomal-recessive disorder characterized by poikiloderma, sparse hair, short stature, and skeletal anomalies. Type 2 RTS, which is defined by the presence of bi-allelic mutations in RECQL4, is characterized by increased cancer susceptibility and skeletal anomalies, whereas the genetic basis of RTS type 1, which is associated with juvenile cataracts, is unknown. We studied ten individuals, from seven families, who had RTS type 1 and identified a deep intronic splicing mutation of the ANAPC1 gene, a component of the anaphase-promoting complex/cyclosome (APC/C), in all affected individuals, either in the homozygous state or in trans with another mutation. Fibroblast studies showed that the intronic mutation causes the activation of a 95 bp pseudoexon, leading to mRNAs with premature termination codons and nonsense-mediated decay, decreased ANAPC1 protein levels, and prolongation of interphase. Interestingly, mice that were heterozygous for a knockout mutation have an increased incidence of cataracts. Our results demonstrate that deficiency in the APC/C is a cause of RTS type 1 and suggest a possible link between the APC/C and RECQL4 helicase because both proteins are involved in DNA repair and replication.

RECQ DNA Helicases and Osteosarcoma.

The RECQ family of DNA helicases is a conserved group of enzymes that plays an important role in maintaining genomic stability. Humans possess five RECQ helicase genes, and mutations in three of them - BLM, WRN, and RECQL4 - are associated with the genetic disorders Bloom syndrome, Werner syndrome, and Rothmund-Thomson syndrome (RTS), respectively. These syndromes share overlapping clinical features, and importantly they are all associated with an increased risk of cancer. Patients with RTS have the highest specific risk of developing osteosarcoma compared to all other cancer predisposition syndromes; therefore, RTS serves as a relevant model to study the pathogenesis and molecular genetics of osteosarcoma. The "tumor suppressor" function of the RECQ helicases continues to be an area of active investigation. This chapter will focus primarily on the known cellular functions of RECQL4 and how these may relate to tumorigenesis, as well as ongoing efforts to understand RECQL4's functions in vivo using animal models. Understanding the RECQ pathways will provide insight into avenues for novel cancer therapies in the future.

Generalized metabolic bone disease and fracture risk in Rothmund-Thomson syndrome.

Rothmund-Thomson syndrome (RTS) is a rare autosomal recessive disorder characterized by poikiloderma, small stature, sparse hair, skeletal abnormalities, increased risk of osteosarcoma, and decreased bone mass. To date, there has not been a comprehensive evaluation of the prevalence and extent of metabolic bone disease in RTS. Furthermore, the mechanisms that result in this phenotype are largely unknown. In this report, we provide a detailed evaluation of 29 individuals with RTS with respect to their metabolic bone status including bone mineral density, calcium kinetics studies, and markers of bone remodeling. We show that individuals with RTS have decreased areal bone mineral density. Additionally, we demonstrate that the presence of pathogenic variants in RECQL4 and low bone mineral density correlate with the history of increased risk of fractures. Using a RECQL4-deficient mouse model that recapitulates skeletal abnormalities seen in individuals with RTS, we demonstrate that generalized skeletal involvement is likely due to decreased osteogenesis. Our findings are clinically relevant as they may help in the risk stratification of patients with RTS and also in the identification of individuals who may benefit from additional surveillance and management of metabolic bone disease.

RECQ DNA helicases and osteosarcoma.

The RECQ family of DNA helicases is a conserved group of enzymes that are important for maintaining genomic integrity. In humans, there are five RECQ helicase genes, and mutations in three of them-BLM, WRN, and RECQL4-are associated with the genetic disorders Bloom syndrome, Werner syndrome, and Rothmund-Thomson syndrome (RTS), respectively. Importantly all three diseases are cancer predisposition syndromes. Patients with RTS are highly and uniquely susceptible to developing osteosarcoma; thus, RTS provides a good model to study the pathogenesis of osteosarcoma. The "tumor suppressor" role of RECQL4 and the other RECQ helicases is an area of active investigation. This chapter reviews what is currently known about the cellular functions of RECQL4 and how these may relate to tumorigenesis, as well as ongoing efforts to understand RECQL4's functions in vivo using animal models. Understanding the RECQ pathways may provide insight into avenues for novel cancer therapies in the future.

Distinguishing between monozygotic twins' blood samples through immune repertoire sequencing.

Monozygotic (MZ) twins with highly similar genomic DNA sequences can not be distinguished by conventional forensic DNA testing. The immune repertoire (IR) reflects an individual's immune history, which is unique between individuals, has been applied to individualized treatment in precision medicine. However, the application of IR in forensic genetics has not been reported to date. In this study, the diversity in the complementary determining region 3 (CDR3) of both the T-cell receptor ß chain (TCRß) and B-cell receptor heavy chain (also known as immunoglobulin heavy chain, IGH) in four pairs of MZ twins were analyzed. The results showed that the amino acid sequences length distribution frequency of TCRß CDR3 had 4-10 differences, and the nucleic acid sequences length distribution frequency of TCRß CDR3 had 2-7 differences between MZ twins. The shared difference of four pairs of MZ twins focused on the length distribution frequency of 34 bp nucleotide sequences in TCRß. By analyzing the usage frequency of V and J genes in TCRß and IGH CDR3 DNA sequence rearrangements, we also found that there were biases between each pair of MZ twins, and the usage frequency of TRBJ2-3 showed common differences between each pair of MZ twins. Furthermore, each pair of MZ twins had its own unique V-J genes combination mode in TCRß and IGH CDR3 DNA sequences. This study, for the first time, suggested that IR can be used as a potential biological marker to distinguish MZ twins.

Complex N-glycans are essential, but core 1 and 2 mucin O-glycans, O-fucose glycans, and NOTCH1 are dispensable, for mammalian spermatogenesis.

To identify roles in spermatogenesis for major subclasses of N- and O-glycans and Notch signaling, male mice carrying floxed C1galt1, Pofut1, Notch1 or Mgat1 alleles and a testis-specific Cre recombinase transgene were generated. T-synthase (C1GALT1) transfers Gal to generate core 1 and core 2 mucin O-glycans; POFUT1 transfers O-fucose to particular epidermal growth factor-like repeats and is essential for canonical Notch signaling; and MGAT1 (GlcNAcT-I) transfers GlcNAc to initiate hybrid and complex N-glycan synthesis. Cre recombinase transgenes driven by various promoters were investigated, including Stra8-iCre expressed in spermatogonia, Sycp1-Cre expressed in spermatocytes, Prm1-Cre expressed in spermatids, and AMH-Cre expressed in Sertoli cells. All Cre transgenes deleted floxed alleles, but efficiencies varied widely. Stra8-iCre was the most effective, deleting floxed Notch1 and Mgat1 alleles with 100% efficiency and floxed C1galt1 and Pofut1 alleles with ~80% efficiency, based on transmission of deleted alleles. Removal of C1galt1, Pofut1, or Notch1 in spermatogonia had no effect on testicular weight, histology, or fertility. However, males in which the synthesis of complex N-glycans was blocked by deletion of Mgat1 in spermatogonia did not produce sperm. Spermatogonia, spermatocytes, and spermatids were generated, but most spermatids formed giant multinucleated cells or symplasts, and apoptosis was increased. Therefore, although core 1 and 2 mucin O-glycans, NOTCH1, POFUT1, O-fucose glycans, and Notch signaling are dispensable, MGAT1 and complex N-glycans are essential for spermatogenesis.

Slc35c2 promotes Notch1 fucosylation and is required for optimal Notch signaling in mammalian cells.

Mammalian Notch receptors require modification by fucose on epidermal growth factor-like (EGF) repeats of their extracellular domain to respond optimally to signal induction by canonical Notch ligands. Inactivation of the Golgi GDP-fucose transporter Slc35c1 in mouse or human does not cause marked defects in Notch signaling during development, and shows milder fucosylation defects than those observed in mice unable to synthesize GDP-fucose, indicating the existence of another mechanism for GDP-fucose transport into the secretory pathway. We show here that fibroblasts from mice or humans lacking Slc35c1 exhibit robust Notch signaling in co-culture signaling assays. A potential candidate for a second GDP-fucose transporter is the related gene Slc35c2. Overexpression of Slc35c2 reduces expression of the fucosylated epitopes Lewis X and sialylated Lewis X in CHO cells, indicating competition with Slc35c1. The fucosylation of a Notch1 EGF repeat fragment that occurs in the endoplasmic reticulum was increased in CHO transfectants overexpressing Slc35c2. In CHO cells with low levels of Slc35c2, both Delta1- and Jagged1-induced Notch signaling were reduced, and the fucosylation of a Notch1 fragment was also decreased. Immunofluorescence microscopy of rat intestinal epithelial cells and HeLa cells, and analysis of rat liver membrane fractions showed that Slc35c2 is primarily colocalized with markers of the cis-Golgi network and endoplasmic reticulum-Golgi intermediate compartment (ERGIC). The combined results suggest that Slc35c2 is either a GDP-fucose transporter that competes with Slc35c1 for GDP-fucose, or a factor that otherwise enhances the fucosylation of Notch and is required for optimal Notch signaling in mammalian cells.

Mix-cultured aerobic denitrifying bacterial communities reduce nitrate: Novel insights in micro-polluted water treatment at lower temperature.

Three mix-cultured aerobic denitrifiers were screened from a source water reservoir and named HE1, HE3 and SU4. Approximately 72.9%, 68.6% and 66.2% of nitrate were effectively removed from basal medium, respectively, after 120 h of cultivation at 8 °C. The nitrogen balance analysis revealed about one-fifth of the initial nitrogen was converted into gaseous denitrification products. According to the results of Biolog, the three microfloras had high metabolic capacity to carbon sources. The dominant genera were Pseudomonas and Paracoccus in these bacterial communities based on nirS gene sequencing. Response surface methodology elucidated that the denitrification rates of identified bacteria reached the maximum under the following optimal parameters: C/N ratio of 7.51-8.34, pH of 8.03-8.09, temperature of 18.03-20.19 °C, and shaking speed of 67.04-120 rpm. All results suggested that screened aerobic denitrifiers could potentially be applied to improve the source water quality at low temperature.

[Effect of Artificial Mixing on Temporal and Spatial Succession of Algae Community Structure in Jinpen Reservoir].

To explore the effect mechanism of the artificial mixing process on the temporal and spatial succession of algae community structure in a water body, this study used water-lifting aerators to induce in-situ artificial mixing of the water body of Jinpen Reservoir, and in-situ spot physical-chemical parameters and algae of the water body of the reservoir were observed during an artificial mixing process. A total of 51 species of 28 genera of 6 families of algae were identified in the water body of the Jinpen Reservoir. The artificial mixing effect of the water-lifting aerators significantly inhibited the growth of algae in the water, and had a significant impact on the community structure. Before activation of the water-lifting aerators, algae were mainly distributed in the surface water body, and Chlorella vulgaris was the dominant species. With the operation of the water-lifting aerators, the algal density of surface water body decreased significantly, and the vertical distribution of the algae density in the water body tended to be uniform. The dominant species tended to succeed in Cyclotella sp. This study used the method of redundancy analysis, combined with critical depth theory and the characteristics of algae growth, to analyze the relationship between the spatial-temporal succession of algae community structure and the changes in the main physical-chemical parameters in Jinpen Reservoir during the artificial process. The analysis results showed that the artificial mixing of the water-lifting aerators mainly affects the temporal and spatial succession of the algae community structure by rapidly destroying the thermal stratification stability of the water body and significantly increasing the water mixing depth.

[Release Mechanisms of Iron and Manganese from Sediments in Jinpen Reservoir].

In response to the annual hypolimnetic anoxia in stratified reservoirs, water-lifting aerators (WLAs) were used in Jinpen Reservoir to supplement the dissolved oxygen in the bottom water and suppress the release of reduced pollutants from sediment. However, due to the influence of geomorphic characteristics at the bottom of the reservoir, there were some differences in the efficiency of artificial mixing and aeration. After the deactivation of WLAs, the dissolved oxygen in the bottom water of some deeper areas was rapidly depleted, resulting in the re-release of pollutants. To explore the release mechanisms and diffusion intensity of iron and manganese during this period, the representative samples in the main reservoir area were collected to measure the distribution of dissolved iron and manganese in the pore water and overlying water and calculate the diffusive flux of dissolved iron and manganese across the sediment-water interface. The results showed that the bottom water of the lower terrain rapidly entered the anaerobic condition after the system was deactivated, resulting in the release of a large amount of dissolved manganese into the overlying water, the maximum concentration of which was 0.42 mg·L-1. However, the bottom water of the higher terrain briefly entered a state of hypoxia, after which the dissolved oxygen concentration increased rapidly, so the dissolved manganese concentration increased moderately to 0.17 mg·L-1. The distribution of iron and manganese in the pore-water-overlying water showed that the dissolved manganese was released more easily into the overlying water than the iron under anaerobic conditions and constant accumulation in the upper sediments and overlying water. However, the release of dissolved iron was not only suppressed by dissolved oxygen but also by other oxidants such as manganese oxide. The diffusion flux of dissolved manganese declined after the system was deactivated. A mass balance calculation demonstrated that the accumulation of dissolved manganese in the anaerobic layer was not only related to the diffusion flux but also to the sedimentation flux and the thickness of the anaerobic layer. Therefore, the biogeochemical cycle of iron and manganese in the anaerobic layer requires further study.

[Characteristics of Organic Matter in Sediments During the Thermal Stratification of a Reservoir and Effects on an Aerobic Denitrifier].

In this study, the relative molecular weight distribution and fluorescent characteristics of the organic matter in sediments during the thermal stratification of a drinking water reservoir were studied. The nitrogen removal, growth performance, and carbon removal ability of an aerobic denitrifier were investigated when the organic matter in sediments was used as a carbon source. The results found that:① during the stratification period in the drinking water reservoir, the organic matter in sediments has a larger proportion of relative molecular mass>100×103. It can be observed that compared with the relative molecular weight distribution in different months, the percentage of macromolecular organic matter in sediments is the lowest in July (44.62%), showing a characteristic of smaller relative molecular weight; ② the organic matter in sediments of the drinking water reservoir was composed of terrestrial humic-like substance component C1 (250 nm, 425 nm), tryptophan and amino acid-like substances component C2 (230 nm/280 nm, 322 nm), and traditional microbial humic-like substances component C3 (250 nm, 340 nm). Component C2 accounted for a higher percentage, and the organic matter in July showed a higher total fluorescence intensity; ③ during the aerobic denitrification process, organic matter in May displayed better characteristics as an electron donor, while organic matter in July exhibited excellent performance as an energy substance and better denitrification characteristics of the strain WGX-9; ④ the aerobic denitrification performance of the strain WGX-9 can be significantly promoted when the organic matter in sediments is a carbon source, compared with natural organic matter, algae organic matter, and actual water of the drinking water reservoir. This study clarifies the characteristics of the organic matter in sediments during the thermal stratification period of the drinking water reservoir and its effect on an aerobic denitrifier. This will provide a scientific basis for the research of nitrogen pollution control in micro-polluted water sources.

[Spatial and Temporal Succession Characteristics of Aerobic Anoxygenic Photosynthesis Bacteria in a Stratified Reservoir].

Aerobic anoxygenic photosynthesis bacteria (AAPB) play a significant role in the material circulation of the hydrosphere, with diverse community structure and unique metabolic functions. To investigate the spatial and temporal succession characteristics of AAPB abundance and community structure in Jinpen Reservoir, a quantitative real-time polymerase chain reaction and Illumina MiSeq high-throughput sequencing technique targeting the pufM gene were applied. Furthermore, redundancy analysis was used to determine the influence of environmental factors on their community structure. The results showed that the AAPB abundance ranged from (6.70±0.43)×103 to (2.69±0.15)×104 copies·mL-1, with the maximum value appearing in October, and decreased with an increase in water depth. Samples were mainly classified into 19 genera (except for the unclassified genus); the most abundant AAPB genera were Bradyrhizobium sp. and Methylobacterium sp., which were affiliated to the α-Proteobacteria, and the proportion of the Bradyrhizobium sp. was highest in November, accounting for more than 60% (except 10 m). Furthermore, Rubrivivax sp., belonging to ß-Proteobacteria, was found to have a low proportion. There was a strong interaction relationship between AAPB genera. For example, Rhodobacter sp. was positively correlated with Rhodovulum sp., while Hydrogenophaga sp. was negatively correlated with Bradyrhizobium sp.. The community structure composition and distribution of AAPB were significantly different, mainly affected by temperature (T), total nitrogen (TN), NO3--N, and light intensity and comprehensively regulated by environmental factors. For instance, T, TN, and total phosphorus had a significant impact on the AAPB community structure of water samples at 0, 5, and 15 m in October, whereas light intensity, pH, DO, and chlorophyll-a were major structuring factors in the AAPB assemblages of water samples at 5 m in December. The results have guiding significance for parsing the spatial and temporal variability of AAPB abundance and diversity in stratified reservoirs, and simultaneously provide a theoretical basis for exploring the driving factors of AAPB population structure.

Canonical Notch signaling is dispensable for early cell fate specifications in mammals.

The canonical Notch signaling pathway mediated by Delta- and Jagged-like Notch ligands determines a variety of cell fates in metazoa. In Caenorhabditis elegans and sea urchins, canonical Notch signaling is essential for different cell fate specifications during early embryogenesis or the formation of endoderm, mesoderm, or ectoderm germ layers. Transcripts of Notch signaling pathway genes are present during mouse blastogenesis, suggesting that the canonical Notch signaling pathway may also function in early mammalian development. To test this directly, we used conditional deletion in oocytes carrying a ZP3Cre recombinase transgene to generate mouse embryos lacking both maternal and zygotic protein O-fucosyltransferase 1, a cell-autonomous and essential component of canonical Notch receptor signaling. Homozygous mutant embryos derived from eggs lacking Pofut1 gene transcripts developed indistinguishably from the wild type until approximately embryonic day 8.0, a postgastrulation stage after the formation of the three germ layers. Thus, in contrast to the case with C. elegans and sea urchins, canonical Notch signaling is not required in mammals for earliest cell fate specifications or for formation of the three germ layers. The use of canonical Notch signaling for early cell fate specifications by lower organisms may represent co-option of a regulatory pathway originally used later in development by all metazoa.

Generation of an induced pluripotent stem cell line from an individual with a heterozygous RECQL4 mutation.

The DNA helicase RECQL4 is known for its roles in DNA replication and repair. RECQL4 mutations cause several genetic disorders including Rothmund-Thomson syndrome (RTS), characterized by developmental defects and predisposition to osteosarcoma. Here we reprogrammed fibroblasts with a heterozygous RECQL4 mutation (c.1878 + 32_1878 + 55del24) to induced pluripotent stem cells (iPSCs). These iPSCs are pluripotent and are able to be differentiated into all three germ layers, providing a novel tool to further interrogate the role of RECQL4 DNA helicase in vitro.

Aging in Rothmund-Thomson syndrome and related RECQL4 genetic disorders.

Rothmund-Thomson Syndrome (RTS) is a rare autosomal recessive disease which manifests several clinical features of accelerated aging. These findings include atrophic skin and pigment changes, alopecia, osteopenia, cataracts, and an increased incidence of cancer for patients carrying RECQL4 germline mutations. Mutations in RECQL4 are responsible for the majority of cases of RTS. RECQL4 belongs to RECQ DNA helicase family which has been shown to participate in many aspects of DNA metabolism. In the past several years, accumulated evidence indicates that RECQL4 is important not only in cancer development but also in the aging process. In this review, based on recent research data, we summarize the common aging findings in RTS patients and propose possible mechanisms to explain the aging features in these patients.

Osteosarcoma: Molecular Pathogenesis and iPSC Modeling.

Rare hereditary disorders provide unequivocal evidence of the importance of genes in human disease pathogenesis. Familial syndromes that predispose to osteosarcomagenesis are invaluable in understanding the underlying genetics of this malignancy. Recently, patient-derived induced pluripotent stem cells (iPSCs) have been successfully utilized to model Li-Fraumeni syndrome (LFS)-associated bone malignancy, demonstrating that iPSCs can serve as an in vitro disease model to elucidate osteosarcoma etiology. We provide here an overview of osteosarcoma predisposition syndromes and review recently established iPSC disease models for these familial syndromes. Merging molecular information gathered from these models with the current knowledge of osteosarcoma biology will help us to gain a deeper understanding of the pathological mechanisms underlying osteosarcomagenesis and will potentially aid in the development of future patient therapies.