ABSTRACT
In the search for novel influenza inhibitors we evaluated 7-fluoro-substituted indoles as bioisosteric replacements for the 7-azaindole scaffold of Pimodivir, a PB2 (polymerase basic protein 2) inhibitor currently in clinical development. Specifically, a 5,7-difluoroindole derivative 11a was identified as a potent and metabolically stable influenza inhibitor. 11a demonstrated a favorable oral pharmacokinetic profile and in vivo efficacy in mice. In addition, it was found that 11a was not at risk of metabolism via aldehyde oxidase, an advantage over previously described inhibitors of this class. The crystal structure of 11a bound to influenza A PB2 cap region is disclosed here and deposited to the PDB.
Subject(s)
Antiviral Agents/chemical synthesis , Antiviral Agents/pharmacology , Drug Design , Indoles/chemical synthesis , Indoles/pharmacology , Viral Proteins/drug effects , A549 Cells , Animals , Antiviral Agents/chemistry , Antiviral Agents/pharmacokinetics , Crystallography, X-Ray , Dogs , Humans , Indoles/chemistry , Indoles/pharmacokinetics , Influenza A Virus, H1N1 Subtype/drug effects , Influenza A Virus, H3N2 Subtype/drug effects , Madin Darby Canine Kidney Cells , Microbial Sensitivity Tests , Molecular StructureABSTRACT
Emergence of drug-resistant bacteria represents a high, unmet medical need, and discovery of new antibacterials acting on new bacterial targets is strongly needed. ATP synthase has been validated as an antibacterial target in Mycobacterium tuberculosis, where its activity can be specifically blocked by the diarylquinoline TMC207. However, potency of TMC207 is restricted to mycobacteria with little or no effect on the growth of other Gram-positive or Gram-negative bacteria. Here, we identify diarylquinolines with activity against key Gram-positive pathogens, significantly extending the antibacterial spectrum of the diarylquinoline class of drugs. These compounds inhibited growth of Staphylococcus aureus in planktonic state as well as in metabolically resting bacteria grown in a biofilm culture. Furthermore, time-kill experiments showed that the selected hits are rapidly bactericidal. Drug-resistant mutations were mapped to the ATP synthase enzyme, and biochemical analysis as well as drug-target interaction studies reveal ATP synthase as a target for these compounds. Moreover, knockdown of the ATP synthase expression strongly suppressed growth of S. aureus, revealing a crucial role of this target in bacterial growth and metabolism. Our data represent a proof of principle for using the diarylquinoline class of antibacterials in key Gram-positive pathogens. Our results suggest that broadening the antibacterial spectrum for this chemical class is possible without drifting off from the target. Development of the diarylquinolines class may represent a promising strategy for combating Gram-positive pathogens.
Subject(s)
ATP Synthetase Complexes/antagonists & inhibitors , Anti-Bacterial Agents/pharmacology , Gram-Positive Bacteria/drug effects , Mitochondria/drug effects , Quinolines/pharmacology , Staphylococcus aureus/drug effects , ATP Synthetase Complexes/genetics , Adenosine Triphosphate/biosynthesis , Amino Acid Sequence , Biofilms/drug effects , Cell Line, Tumor , Drug Resistance, Bacterial/genetics , Gram-Positive Bacteria/growth & development , HeLa Cells , Humans , Microbial Sensitivity Tests , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/growth & development , Quinolines/chemistry , Quinolines/toxicity , Sequence Alignment , Staphylococcus aureus/growth & developmentABSTRACT
LRP5 was recently confirmed as an important susceptibility gene for osteoporosis. Our objective was to evaluate the effect of DKK1 polymorphisms on bone mineral density (BMD), hip geometry, and bone turnover. DKK1 is a secreted protein that binds to LRP5/6 receptors and inhibits canonical Wnt signaling. Using HapMap, we selected three SNPs covering the genetic variation in a 13.53-kb region comprising DKK1. The Odense Androgen Study is a population-based study comprising 783 Caucasian men aged 20-29 years. BMD and hip structural parameters were available for study. Bone turnover markers were used as a secondary end point. All analyses were repeated after adjusting for covariables and in subgroups according to physical activity. We found no significant association between DKK1 and BMD or markers of bone turnover; however, a significant association (P = 0.012) was found for rs1569198 with hip axis length (HAL), independent of BMD and height. Moreover, the association seemed to be driven by the non-sedentary subgroup (P = 0.004). Haplotype analysis further confirmed the association of rs1569198 with HAL. Furthermore, we obtained indications for interaction between DKK1 and LRP5 genotypes for different hip geometry parameters. As almost all variance within the DKK1 gene was covered, we conclude that common variation in this gene does not markedly influence BMD or bone turnover markers in young men. In this population, however, a common SNP in DKK1 does have a significant effect on HAL, implying a possible effect on hip fracture risk in the general population. This finding could be of interest but needs replication in independent populations.
Subject(s)
Bone Density/genetics , Bone Remodeling/genetics , Hip/anatomy & histology , Intercellular Signaling Peptides and Proteins/genetics , Adult , Biomarkers/analysis , Biomarkers/metabolism , Body Weights and Measures , Cohort Studies , Denmark , Epistasis, Genetic , Genetic Association Studies , Genetic Predisposition to Disease , Genetic Variation/physiology , Hip Fractures/genetics , Humans , Linkage Disequilibrium , Male , Polymorphism, Single Nucleotide/physiology , Young AdultABSTRACT
Recently, Brinster et al. suggested that type II fatty-acid biosynthesis (FASII) is not a suitable antibacterial target for Gram-positive pathogens because they use fatty acids directly from host serum rather than de novo synthesis. Their findings, if confirmed, are relevant for further scientific and financial investments in the development of new drugs targeting FASII. We present here in vitro and in vivo data demonstrating that their observations do not hold for Staphylococcus aureus, a major Gram-positive pathogen causing several human infections. The observed differences among Gram-positive pathogens in FASII reflects heterogeneity either in fatty-acid synthesis or in the capacity for fatty-acid uptake from the environment.
Subject(s)
Fatty Acids/biosynthesis , Staphylococcus aureus/metabolism , Animals , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Bacteremia/drug therapy , Bacteremia/microbiology , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Fatty Acids/metabolism , Fatty Acids/pharmacology , Host-Pathogen Interactions/drug effects , Humans , Mice , Reproducibility of Results , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics , Staphylococcus aureus/pathogenicity , Triclosan/pharmacologyABSTRACT
Sclerosteosis is a severe, rare, autosomal recessive bone condition that is characterized by a progressive craniotubular hyperostosis. The main features are a significant sclerosis of the long bones, ribs, pelvis, and skull, leading to facial distortion and entrapment of cranial nerves. Clinical features include a tall stature, nail dysplasia, cutaneous syndactyly of some fingers, and raised intracranial pressure. The sclerosteosis gene has been mapped to chromosome 17q12-21 and is currently known as the SOST gene encoding the sclerostin protein. Here, we report on one familial and one isolated case of Brazilian origin with the clinical and molecular diagnosis of sclerosteosis. The radiological and clinical features are described, and the diagnosis of sclerosteosis was confirmed in both cases by mutation analysis of the SOST gene showing a homozygous nonsense mutation (Trp124X) in the two patients. We reported this mutation previously in other sclerosteosis patients from a consanguineous Brazilian family. Interestingly, all three families were from the same state in Brazil, but they denied familial relationship. These patients confirm the clinical picture as found in other cases with a loss of function mutation in the SOST gene.
Subject(s)
Bone Morphogenetic Proteins/genetics , Codon, Nonsense , Genetic Markers/genetics , Hyperostosis/genetics , Osteosclerosis/genetics , Adaptor Proteins, Signal Transducing , Adult , Brazil , Child , Female , Homozygote , Humans , Hyperostosis/diagnostic imaging , Hyperostosis/pathology , Male , Osteosclerosis/diagnostic imaging , Osteosclerosis/pathology , Radiography , Young AdultABSTRACT
An estimated one-third of the world population is latently infected with Mycobacterium tuberculosis. These nonreplicating, dormant bacilli are tolerant to conventional anti-tuberculosis drugs, such as isoniazid. We recently identified diarylquinoline R207910 (also called TMC207) as an inhibitor of ATP synthase with a remarkable activity against replicating mycobacteria. In the present study, we show that R207910 kills dormant bacilli as effectively as aerobically grown bacilli with the same target specificity. Despite a transcriptional down-regulation of the ATP synthase operon and significantly lower cellular ATP levels, we show that dormant mycobacteria do possess residual ATP synthase enzymatic activity. This activity is blocked by nanomolar concentrations of R207910, thereby further reducing ATP levels and causing a pronounced bactericidal effect. We conclude that this residual ATP synthase activity is indispensable for the survival of dormant mycobacteria, making it a promising drug target to tackle dormant infections. The unique dual bactericidal activity of diarylquinolines on dormant as well as replicating bacterial subpopulations distinguishes them entirely from the current anti-tuberculosis drugs and underlines the potential of R207910 to shorten tuberculosis treatment.
Subject(s)
Adenosine Triphosphate/chemistry , Gene Expression Regulation, Bacterial , Homeostasis , Mycobacterium/metabolism , Quinolines/pharmacology , Antitubercular Agents/pharmacology , Mitochondrial Proton-Translocating ATPases/chemistry , Models, Biological , Mycobacterium bovis/drug effects , Mycobacterium bovis/metabolism , Mycobacterium smegmatis/drug effects , Mycobacterium smegmatis/metabolism , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/metabolism , Nitric Oxide/chemistry , Oxygen/chemistry , RNA, Messenger/metabolism , Time FactorsABSTRACT
Craniotubular hyperostosis are a group of high bone mass disorders related to mutations in the LRP5 and SOST genes, although other causative genes remain to be identified. Little is known about the bone turnover and the response to T3 or glucocorticoids in these patients. We describe a patient with craniotubular hyperostosis, including mutation analyses of the LRP5, SOST, DKK1 and KRM1 genes. We also studied bone turnover and bone mineral density (BMD), before and after a trial with T3 (75 microg/d for 28 weeks) and T3 and prednisone (T3 100 microg/d for 2 weeks, followed by 10 weeks on prednisone 10 mg/d, and a final 2 weeks period off of medicactions, completing 3 cycles in 42 weeks. Mutation analysis of the complete coding region and flanking highly conserved sequences of SOST, evaluation of the presence of the 52-kb deletion associated with Van Buchem disease in Dutch patients and mutation analysis of exons 2-4 of LRP5, and the coding regions of DKK1 and KRM1 did not reveal any disease-causing mutations. A baseline 5 to 7 fold increase in osteocalcin and in deoxypiridinoline was detected. After 4 weeks on 75 microg/d of T3, osteocalcin decreased 36%, but at week 28, it returned to basal. Deoxypiridinoline did not change. After the first cycle on T3 and prednisone, osteocalcin decreased 72%, and at the end of the third cycle it remained 44% below basal value. Deoxypiridinoline was stable and high during the three cycles; no changes in BMD were observed. As we failed to identify any disease-causing mutations in our patient with craniotubular hyperostosis, we suggest that another gene must be involved in the pathogenesis of his condition. This study provides additional data about the high bone turnover described in craniotubular hyperostosis, and also suggests an abnormal response to T3 excess in this condition.
Subject(s)
Bone and Bones/metabolism , Hyperostosis/drug therapy , Hyperostosis/genetics , Prednisone/therapeutic use , Skull/pathology , Triiodothyronine/therapeutic use , Adult , Bone Density/drug effects , Bone and Bones/drug effects , Child, Preschool , Hand/diagnostic imaging , Humans , Male , Prednisone/pharmacology , Radiography , Triiodothyronine/pharmacologyABSTRACT
Low-density lipoprotein receptor-related protein 5 (LRP5), a Wnt coreceptor, plays an important role in bone metabolism as loss-of-function and gain-of-function mutations in LRP5 result in the autosomal recessive osteoporosis-pseudoglioma syndrome and autosomal dominant high-bone mass (HBM) phenotypes, respectively. Prior studies suggested that the presence of HBM-associated LRP5 mutations results in decreased antagonism of LRP5-mediated Wnt signaling. In the present study, we investigated six different HBM-LRP5 mutations and confirm that neither Dickkopf1 (DKK1) nor sclerostin efficiently inhibits HBM-LRP5 signaling. In addition, when coexpressed, DKK1 and sclerostin do not inhibit HBM-LRP5 mutants better than either inhibitor by itself. Also, DKK1 and sclerostin do not simultaneously bind to wild-type LRP5, and DKK1 is able to displace sclerostin from previously formed sclerostin-LRP5 complexes. In conclusion, our results indicate that DKK1 and sclerostin are independent, and not synergistic, regulators of LRP5 signaling and that the function of each is impaired by HBM-LRP5 mutations.
Subject(s)
Bone Morphogenetic Proteins/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , LDL-Receptor Related Proteins/metabolism , Adaptor Proteins, Signal Transducing , Animals , Cell Line , Culture Media, Conditioned/chemistry , Genetic Markers , Humans , LDL-Receptor Related Proteins/genetics , Low Density Lipoprotein Receptor-Related Protein-5 , Mice , Protein Binding , Signal TransductionABSTRACT
A few years ago, human genetic studies provided compelling evidence that the low-density lipoprotein receptor-related protein 5 (LRP5) is involved in the regulation of bone homeostasis because pathogenic LRP5 mutations were found in monogenic conditions with abnormal bone density. On the one hand, the osteoporosis pseudoglioma syndrome results from loss of function of LRP5, whereas on the other hand, gain-of-function mutations in LRP5 cause conditions with an increased bone density. On the molecular level, these types of mutations result in disturbed (respectively, decreased and increased) canonical Wnt signaling, an important metabolic pathway in osteoblasts during embryonic and postnatal osteogenesis. This signaling cascade is activated by binding of Wnt ligand to the Frizzled/LRP5 receptor complex. In addition to the involvement of LRP5 in conditions with extreme bone phenotypes, the genetic profile of this gene has also been shown to contribute to the determination of bone density in the general population. Quite a number of studies already demonstrated that common polymorphic variants in LRP5 are associated with bone mineral density and consequently osteoporosis, a multifactorial trait with low bone mass and porous bone structure. These genetic studies together with results obtained from in vitro and in vivo studies emphasize the importance of LRP5 and canonical Wnt signaling in the regulation of bone homeostasis. Therefore, unraveling the exact mechanisms of this signaling cascade has become an important area in bone research. This review focuses on the genetics of LRP5 and summarizes the findings on monogenic bone conditions as well as the current knowledge of its involvement in the pathogenesis of osteoporosis.
Subject(s)
Bone and Bones/metabolism , LDL-Receptor Related Proteins/genetics , LDL-Receptor Related Proteins/physiology , Bone Density/genetics , Humans , Low Density Lipoprotein Receptor-Related Protein-5 , Models, Biological , Mutation , Osteoporosis/geneticsSubject(s)
Bone Morphogenetic Proteins/genetics , Deafness/genetics , Foot Bones/abnormalities , Genes, Recessive , Hand Bones/abnormalities , Ketoglutarate Dehydrogenase Complex/genetics , Bone Morphogenetic Protein 4 , Deafness/enzymology , Female , Foot Bones/enzymology , Hand Bones/enzymology , Humans , Infant , Infant, Newborn , Male , Pedigree , Siblings , SyndromeABSTRACT
UNLABELLED: We found a novel heterozygous missense mutation (M282V) in the LRP5 gene in a patient with a high bone mass phenotype. In vitro studies suggest that a reduced antagonistic effect of DKK1 on canonical Wnt signaling contributes to the molecular effect of this mutation and its pathogenic consequence. INTRODUCTION: Gain-of-function mutations in the gene encoding LDL receptor-related protein 5 (LRP5) cause high bone mass. Recent studies revealed that a reduced inhibition of canonical Wnt signaling by Dickkopf 1 (DKK1) contributes to the pathophysiology of this disease phenotype. MATERIALS AND METHODS: We report on a 55-yr-old female patient with a high bone mass phenotype. Sequencing of exons 2-4 of the LRP5 gene was carried out to screen for disease-associated mutations in genomic DNA of the patient. The effect of the identified mutation on LRP5 membrane trafficking was studied by immunoblotting of a truncated form of LRP5. Additionally, Wnt signal activation in the absence and presence of DKK1 was assessed using a TCF4-based reporter gene assay in Saos-2 cells. RESULTS: Our patient presents with dense bones (Z-scores > +6), and radiographic examination showed a generalized thickening of the skeleton. BMD at the hip and lumbar spine significantly decreased through the passage to menopause, indicating no protection to bone loss. Further clinical evaluation revealed torus palatinus. Mutation analysis showed the presence of a novel heterozygous missense variant (844A-->G; M282V) in LRP5, located in the first beta-propeller domain of the extracellular portion. Although protein secretion seemed to be impaired, this mutant was able to transduce Wnt signals at levels comparable with wildtype LRP5. We additionally observed a less efficient inhibition of canonical Wnt signaling by DKK1. CONCLUSIONS: Like all high BMD-associated gain-of-function LRP5 mutations described thus far, the M282V variant affects an amino acid located in the first beta-propeller domain, underlining the functional importance of this region in the pathophysiology of these conditions. This mutation most likely alters a region important for LRP5 modulation by DKK.
Subject(s)
Bone Density/genetics , Bone Diseases/genetics , Intercellular Signaling Peptides and Proteins/metabolism , LDL-Receptor Related Proteins/genetics , Mutation, Missense , Signal Transduction/genetics , Wnt Proteins/metabolism , Amino Acid Substitution , Bone Diseases/diagnostic imaging , Bone Diseases/metabolism , Bone Diseases/physiopathology , Cell Line , Cell Membrane/genetics , Cell Membrane/metabolism , DNA Mutational Analysis , Female , Humans , LDL-Receptor Related Proteins/metabolism , Low Density Lipoprotein Receptor-Related Protein-5 , Middle Aged , Phenotype , Protein Structure, Tertiary/genetics , Protein Transport/genetics , RadiographyABSTRACT
UNLABELLED: We studied whether the LRP5 gene contributes to the clinical phenotype of IO in men. Mutation analysis in 66 IO men revealed a range of sequence variants, of which two missense variants were shown to be of functional relevance. INTRODUCTION: Mutations in the LDL receptor-related protein 5 (LRP5) gene have been associated with extreme bone phenotypes, which makes LRP5 a plausible candidate gene for idiopathic osteoporosis (IO). MATERIALS AND METHODS: In 66 men with IO, all 23 exons and exon-intron boundaries of the LRP5 gene were screened for mutations, and functional analyses were performed for those that were putatively involved in the phenotype. RESULTS: Mutation analysis in the IO probands revealed five missense mutations, of which 1067C>T (S356L), 1364C>T (S455L), and 4609G>A (A1537T) were of potential functional significance because they were located in highly conserved regions of LRP5 and not found in a control panel. Segregation analysis in the respective families could not exclude their possible causality for IO. Furthermore, functional analyses clearly showed an inhibitory effect of mutations 1067C>T and 1364C>T on Wnt signal transduction. These effects are most likely caused by impaired LRP5 synthesis in the case of 1067C>T and failure of protein trafficking to the cell surface for 1364C>T. CONCLUSIONS: For 2 of 66 IO probands, a mutation in the LRP5 gene with proven functionality was found. The findings indicate that carrying an LRP5 mutation is a risk factor for IO, but that overall, IO in men is infrequently underlied by such a mutation.
Subject(s)
LDL-Receptor Related Proteins/genetics , Mutation, Missense/genetics , Osteoporosis/genetics , Adult , Aged , Amino Acid Sequence , Bone Density/genetics , Bone Diseases, Metabolic/genetics , Cell Line , Culture Media, Conditioned/metabolism , Exons/genetics , Gene Frequency , Humans , Introns/genetics , LDL-Receptor Related Proteins/metabolism , Low Density Lipoprotein Receptor-Related Protein-5 , Male , Middle Aged , Molecular Sequence Data , Osteoporosis/etiology , Pedigree , Polymorphism, Genetic/genetics , Protein Transport/genetics , Sequence Homology, Amino Acid , TCF Transcription Factors/genetics , TCF Transcription Factors/metabolism , Transcription Factor 7-Like 2 Protein , Transfection , Wnt1 Protein/geneticsABSTRACT
Gain-of-function mutations in LRP5 have been shown to cause high BMD disorders showing variable expression of some clinical symptoms, including torus palatinus and neurological complications. In an extended family, we were able to add craniosynostosis and developmental delay to the clinical spectrum associated with LRP5 mutations. We report on an extended four-generation family with 13 affected individuals (7 men and 6 women) in which an autosomal dominant type of osteosclerosis segregates. Osteosclerosis was most pronounced in the cranial base and calvarium, starting in early childhood with variable expression and a progressive character. Craniosynostosis at an early age was reported in four affected family members (two males and two females). The patients also presented with dysmorphic features (macrocephaly, brachycephaly, wide and high forehead, hypertelorism, prominent cheekbones, prominent jaw). They have normal height and proportions. Neurological complications like entrapment of cranial nerves resulting in optical nerve atrophy, hearing loss, and facial palsy were reported in two individuals. A mild developmental delay was reported in three affected individuals. None of the patients have torus palatinus, increased rate of fractures, osteomyelitis, hepatosplenomegaly, or pancytopenia. A missense mutation 640G-->A (A214T) in the low-density lipoprotein receptor-related protein 5 (LRP5) gene was found in all affected individuals analyzed, including cases in whom craniosynostosis, a mild developmental delay, and/or macrocephaly is observed. To our knowledge, this is the first report in the literature of patients presenting with autosomal dominant osteosclerosis in whom a variable expression of craniosynostosis, macrocephaly, and mild developmental delay is observed, which is most likely associated with a mutation in the LRP5 gene. These phenotypes can therefore be added to the clinical spectrum of LRP5-associated bone disorders.
Subject(s)
Craniosynostoses/genetics , LDL-Receptor Related Proteins/genetics , Mutation, Missense , Osteosclerosis/genetics , Adult , DNA Mutational Analysis , Female , Humans , Low Density Lipoprotein Receptor-Related Protein-5 , Male , Middle Aged , Nuclear Proteins/genetics , Osteosclerosis/complications , Pedigree , Phenotype , Protein-Tyrosine Kinases/genetics , Radiography , Receptor Protein-Tyrosine Kinases/genetics , Receptor, Fibroblast Growth Factor, Type 2 , Receptor, Fibroblast Growth Factor, Type 3 , Receptors, Fibroblast Growth Factor/genetics , Skull/diagnostic imaging , Skull/pathology , Transcription Factors/genetics , Twist-Related Protein 1ABSTRACT
In this study, a brother and sister of German origin are described with a possible diagnosis of van Buchem disease, a rare autosomal recessive sclerosing bone dysplasia characterized by a generalized hyperostosis of the skeleton mainly affecting the cranial bones. Clinically, patients suffer from cranial nerve entrapment potentially resulting in facial paresis, hearing disturbances, and visual loss. The radiological picture of van Buchem disease closely resembles sclerosteosis, although in the latter patients, syndactyly, tall stature, and raised intracranial pressure are frequently observed, allowing a differential diagnosis with van Buchem disease. Previous molecular studies demonstrated homozygous loss-of-function mutations in the SOST gene in sclerosteosis patients while a chromosomal rearrangement creating a 52-kb deletion downstream of this gene was found in Dutch patients with van Buchem disease. This deletion most likely suppresses SOST expression. Sclerostin, the SOST gene product, has been shown to play a role in bone metabolism. The two siblings reported here were evaluated at the molecular level by carrying out a mutation analysis of the SOST gene. This resulted in the identification of a novel putative disease-causing splice site mutation (IVS1 + 1 G-->C) homozygously present in both siblings.
Subject(s)
Bone Morphogenetic Proteins/genetics , Genetic Markers/genetics , Hyperostosis/genetics , Siblings , Adaptor Proteins, Signal Transducing , Adult , Bone Diseases/genetics , Bone Diseases/pathology , Bone and Bones/pathology , DNA/analysis , DNA/isolation & purification , Facies , Female , Homozygote , Humans , Hyperostosis/pathology , Leukocytes, Mononuclear/chemistry , Male , Point Mutation/genetics , Polymerase Chain Reaction , RNA Splice Sites/geneticsABSTRACT
Van Buchem disease is an autosomal recessive disease characterized by overgrowth of the skeleton. In a group of Dutch patients the disease is thought to be due to a 52-kb deletion that results in decreased expression of the SOST gene. To further characterize the disease, the morphology of the metacarpals of six adult subjects and two juveniles with Van Buchem disease were measured on hand x-rays along with nine normal adults and nine adult carriers of the disease. Serum bone formation markers, alkaline phosphatase, type I procollagen peptide, and osteocalcin, and the urinary bone resorption marker, cross-linked N-telopeptide, were determined. Van Buchem patients had increased metacarpal outer diameter, inner diameter, cortical thickness, and bone mineral density. Calculated bone volume and derived polar moment of inertia were markedly elevated (elevations of 158 +/- 33% and 497 +/- 95%, respectively) consistent with increased bone strength. Serum procollagen peptide and osteocalcin were significantly higher in Van Buchem patients. Urinary cross-linked N-telopeptide was significantly elevated in Van Buchem patients. None of these changes was found in Van Buchem carriers. These observations indicate that decreased expression of the SOST gene can lead to increased bone formation and to stronger bones.
Subject(s)
Bone Density , Hyperostosis, Cortical, Congenital/genetics , Hyperostosis, Cortical, Congenital/physiopathology , Osteogenesis , Adolescent , Adult , Aged , Biomarkers/blood , Biomarkers/urine , Bone Resorption/metabolism , Case-Control Studies , Child , Collagen/urine , Collagen Type I , Female , Genes, Recessive , Hand/diagnostic imaging , Heterozygote , Humans , Hyperostosis, Cortical, Congenital/diagnostic imaging , Male , Metacarpus/diagnostic imaging , Middle Aged , Peptides/urine , RadiographyABSTRACT
OBJECTIVE: The purpose of this study was to evaluate the lifetime evolution of the radioclinical features in a large family with van Buchem disease. DESIGN AND PATIENTS: The study population included 13 patients, ranging between 6 and 69 years. The evolution of the clinical features has been assessed by retrospective analysis of the clinical records of the patients. The age-related evolution of the cortical hyperostosis and defective modeling at the tubular bones was evaluated by morphometric analysis of hand films in 9 patients, compared with 9 control individuals. Progression of sclerosis of the craniofacial bones was evaluated by analysis of the skull radiographs of eleven van Buchem patients, taken at different age. RESULTS AND CONCLUSIONS: Radioclinical features, including sclerosis of the cranial and tubular bones and cranial nerve deficit, become more prominent in older patients. Defective modeling of tubular bones, cortical thickness and medullary width progress with age. Radioclinical abnormalities of van Buchem patients become more prominent in older patients, which suggests that the van Buchem gene is very actively involved in bone metabolism throughout life. Morphometric analysis of the plain films supports the hypothesis that the physiological function of the van Buchem gene is to inhibit bone formation and possibly to regulate bone remodeling.
Subject(s)
Bone Diseases, Developmental/genetics , Bone Diseases, Developmental/pathology , Adolescent , Adult , Age Factors , Aged , Bone Diseases, Developmental/diagnostic imaging , Camurati-Engelmann Syndrome/diagnostic imaging , Camurati-Engelmann Syndrome/genetics , Camurati-Engelmann Syndrome/pathology , Child , Craniofacial Abnormalities/diagnostic imaging , Craniofacial Abnormalities/genetics , Craniofacial Abnormalities/pathology , Diaphyses , Facial Nerve Diseases/diagnostic imaging , Facial Nerve Diseases/genetics , Facial Nerve Diseases/pathology , Family Health , Female , Genetic Predisposition to Disease/genetics , Hearing Loss, Conductive/diagnostic imaging , Hearing Loss, Conductive/genetics , Hearing Loss, Conductive/pathology , Hearing Loss, Sensorineural/diagnostic imaging , Hearing Loss, Sensorineural/genetics , Hearing Loss, Sensorineural/pathology , Humans , Hyperostosis, Cortical, Congenital/diagnostic imaging , Hyperostosis, Cortical, Congenital/genetics , Hyperostosis, Cortical, Congenital/pathology , Male , Metacarpus/diagnostic imaging , Metacarpus/pathology , Middle Aged , Netherlands , Observer Variation , Pedigree , Retrospective Studies , Sclerosis , Skull/diagnostic imaging , Skull/pathology , Statistics as Topic , Tomography, X-Ray ComputedABSTRACT
The transforming growth factor-beta (TGF-beta) superfamily contains a variety of growth factors which all share common sequence elements and structural motifs. These proteins are known to exert a wide spectrum of biological responses on a large variety of cell types in both vertebrates and invertebrates. Many of them have important functions during embryonic development in pattern formation and tissue specification, and in adult tissues, they are involved in processes such as wound healing, bone repair, and bone remodeling. The family is divided into two general branches: the BMP/GDF and the TGF-beta/Activin/Nodal branches, whose members have diverse, often complementary effects. It is obvious that an orchestered regulation of different actions of these proteins is necessary for proper functioning. The TGF-beta family members act by binding extracellularly to a complex of serine/threonine kinase receptors, which consequently activate Smad molecules by phosphorylation. These Smads translocate to the nucleus, where they modulate transcription of specific genes. Three levels by which this signaling pathway is regulated could be distinguished. First, a control mechanism exists in the intracellular space, where inhibitory Smads and Smurfs prevent further signaling and activation of target genes. Second, at the membrane site, the pseudoreceptor BAMBI/Nma is able to inhibit further signaling within the cells. Finally, a range of extracellular mediators are identified which modulate the functioning of members of the TGF-beta superfamily. Here, we review the insights in the extracellular regulation of members of the BMP subfamily of secreted growth factors with a major emphasis on vertebrate BMP modulation.