The early evolutionary landscape of osteosarcoma provides clues for targeted treatment strategies

Osteosarcomas are aggressive primary tumors of bone that are typically detected in locally advanced stages; however, which genetic mutations drive the cancer before its clinical detection remain unknown. To identify these events, we performed longitudinal genome-sequencing analysis of 12 patients with metastatic or refractory osteosarcoma. Phylogenetic and molecular clock analyses were carried out next to identify actionable mutations, and these were validated by integrating data from additional 153 osteosarcomas and pre-existing functional evidence from mouse PDX models. We found that the earliest and thus clinically most promising mutations affect the cell cycle G1 transition, which is guarded by cyclins D3, E1, and cyclin-dependent kinases 2, 4, and 6. Cell cycle G1 alterations originate no more than a year before the primary tumor is clinically detected and occur in >90% and 50% of patients of the discovery and validation cohorts, respectively. In comparison, other cancer driver mutations could be acquired at any evolutionary stage and often do not become pervasive. Consequently, our data support that the repertoire of actionable mutations present in every osteosarcoma cell is largely limited to cell cycle G1 mutations. Since they occur in mutually exclusive combinations favoring either CDK2 or CDK4/6 pathway activation, we propose a new genomically-based algorithm to direct patients to correct clinical trial options

Convergent Evolution of Copy Number Alterations in Multi-Centric Hepatocellular Carcinoma

In the recent years, new molecular methods have been proposed to discriminate multicentric hepatocellular carcinomas (HCC) from intrahepatic metastases. Some of these methods utilize sequencing data to assess similarities between cancer genomes, whilst other achieved the same results with transcriptome and methylome data. Here, we attempt to classify two HCC patients with multi-centric disease using the recall-rates of somatic mutations but find that difficult because their tumors share some chromosome-scale copy-number alterations (CNAs) but little-to-no single-nucleotide variants. To resolve the apparent conundrum, we apply a phasing strategy to test if those shared CNAs are identical by descent. Our findings suggest that the conflicting alterations occur on different homologous chromosomes, which argues for multi-centric origin of respective HCCs

Roles of Gut Microbiome in Bone Homeostasis and Its Relationship with Bone-Related Diseases

The extended microbial genome—the gut microbiome (GM)—plays a significant role in host health and disease. It is able to influence a number of physiological functions. During dysbiosis, GM is associated with the development of various chronic diseases with impaired bone quality. In general, GM is important for bone homeostasis and can affect it via several mechanisms. This review describes the roles of GM in bone homeostasis through influencing the immune and endocrine functions, short-chain fatty acids production, calcium absorption and the gut–brain axis. The relationship between GM composition and several bone-related diseases, specifically osteoporosis, osteoarthritis, rheumatoid arthritis, diabetes mellitus, obesity and bone cancer, is also highlighted and summarized. GM manipulation may become a future adjuvant therapy in the prevention of many chronic diseases. Therefore, the beneficial effects of probiotic therapy to improve the health status of individuals with aforementioned diseases are provided, but further studies are needed to clearly confirm its effectiveness. Recent evidence suggests that GM is responsible for direct and indirect effects on drug efficacy. Accordingly, various GM alterations and interactions related to the treatment of bone-related diseases are mentioned as well

The impact of cadmium exposure on cell parameters, expression of specific genes and mineralization in cultivated human osteoblasts

Cadmium (Cd) is a widespread environmental pollutant which negatively affects bone health. The aim of this study was to investigate the impact of Cd exposure on cell parameters, expression of specific genes and mineralization in cultivated human osteoblasts. Osteoblasts were incubated without (control group) or with CdCl2 at final Cd concentrations of 0.1, 2, 5, 10, and 50 μM for 2, 4, 6 or 14 days. Cell viability, morphological changes, alkaline phosphatase activity, mineralization, and an expression of 10 genes associated with osteoblast-specific pathways, oxidative stress and cell death were determined. Osteoblast viability decreased at the highest Cd concentrations (5, 10 and 50 μM) after 4 days of treatment. Our findings specified the threshold of Cd exposure with morphological alterations to 5 μM Cd following 4 days of cultivation. Morphometric measurements revealed significantly decreased size of Cd-treated osteoblasts when compared to the control. The alkaline phosphatase activity was dose-dependent with a reduction at 5 μM Cd after 2 days. Cd concentration of 5 μM significantly down-regulated the expression of COL1A1, ALPL, BGLAP, and GPX1 genes. On the contrary, BAX and SOD1 genes were over-expressed at 0.1, 2, 5 μM Cd and TNFSF11 gene transcripts showed an increase at 0.1-2 μM Cd. No significant impact of Cd exposure on transcription levels of WNT5A, RUNX2, and CASP1 genes was detected. Cd at 0.1 μM stimulated the mineralization, while the dose of 1 μM Cd had no effect. In conclusion, Cd exposure had adverse impact on human osteoblasts viability, morphology and function. On molecular level, it was associated with bone matrix protein synthesis, bone remodelling, cell death, and response to oxidative stress. The lowest dose of Cd had the stimulatory effect on bone mineralization

Synthesis of Novel Biologically Active Proflavine Ureas Designed on the Basis of Predicted Entropy Changes

A novel series of proflavine ureas, derivatives 11a–11i, were synthesized on the basis of molecular modeling design studies. The structure of the novel ureas was obtained from the pharmacological model, the parameters of which were determined from studies of the structure-activity relationship of previously prepared proflavine ureas bearing n-alkyl chains. The lipophilicity (LogP) and the changes in the standard entropy (ΔS°) of the urea models, the input parameters of the pharmacological model, were determined using quantum mechanics and cheminformatics. The anticancer activity of the synthesized derivatives was evaluated against NCI-60 human cancer cell lines. The urea derivatives azepyl 11b, phenyl 11c and phenylethyl 11f displayed the highest levels of anticancer activity, although the results were only a slight improvement over the hexyl urea, derivative 11j, which was reported in a previous publication. Several of the novel urea derivatives displayed GI50 values against the HCT-116 cancer cell line, which suggest the cytostatic effect of the compounds azepyl 11b–0.44 μM, phenyl 11c–0.23 μM, phenylethyl 11f–0.35 μM and hexyl 11j–0.36 μM. In contrast, the novel urea derivatives 11b, 11c and 11f exhibited levels of cytotoxicity three orders of magnitude lower than that of hexyl urea 11j or amsacrine

Adverse changes in cortical and trabecular bone compartments of the femur in rats with adjuvant-induced arthritis after one remodelling cycle

It is widely known that rheumatoid arthritis (RA) is associated with articular bone damage. However, there is still not enough information on whether the inflammatory process can deteriorate bone microstructure outside the joint as well. Furthermore, the impact of RA on the microscopic structure of cortical and trabecular bone, including parameters of bone microarchitecture, strength, and geometry after one remodelling cycle, has not been determined, yet. Therefore, this study investigated possible alterations in both cortical and trabecular bone compartments of the femur in a rat model of adjuvant-induced arthritis (AA) 28 days post disease induction. AA was generally evoked by a single intradermal injection ofsuspension of heat-inactivated Mycobacterium butyricumin incomplete Freund’s adjuvant. We have found that AA resulted in inflammation as evidenced by increased hind paw swelling, decreased levels of circulating albumin, and elevated levels of nitrite/nitrate, interleukin-1β. Detrimental changes in examined bone parameters related to microarchitecture, strength, and geometry were revealed in AA rats. Overall, AA was associated with bone loss, decreased bone mineral density in bothcortical and trabecular bone compartments, as well as reduced mechanical competence, and more intense vascularization in the cortical bone. According to our results, AA-related inflammation caused structural degradation of cortical and trabecular bone quality, as well as mechanical weakness in the femoral diaphysis leading to bone fragility after only one remodelling cycle. The findings focused on the femoral diaphysis, which is located outside the joint, are the first in this field of research

Bee Bread Can Alleviate Lipid Abnormalities and Impaired Bone Morphology in Obese Zucker Diabetic Rats

This study examined for the first time whether bee bread (BB, consisting of monofloral rape bee pollen) could alleviate lipid derangements and reduced bone quality in Zucker diabetic fatty (ZDF) rats, which are considered an appropriate animal model for type 2 diabetes mellitus (T2DM) investigation. Adult ZDF rats were segregated into four groups: lean non-diabetic rats (L group), obese diabetic rats untreated (C group), and those treated with the BB at two doses (500 and 700 mg/kg body weight, respectively, B1 and B2 groups) for 10 weeks. Significantly reduced levels of total cholesterol and triglyceride were recorded in the B2 group versus the C group. In both BB-treated groups, significantly increased relative volume of trabecular bone and trabecular thickness, enhanced density of secondary osteons, accelerated periosteal bone apposition, and improved blood flow were observed. A positive effect of higher dose of BB on femoral weight and cortical bone thickness was also demonstrated. Our results suggest a promising potential of BB to ameliorate T2DM-related complications associated with lipid and bone damages

Subacute exposure to amygdalin influences compact bone remodeling of rabbits

Amygdalin is most commonly occurring cyanogenic glycoside. It is found in seeds of many plant species. Our study was aimed to reveal whether pure intramuscularly injected amygdalin or apricot seeds peroral exposure cause changes in bone microstructure of rabbits. Twenty clinically healthy 5 months-old male rabbits were segregated into five groups. Animals from groups A1 and A2 were intramuscularly injected with amygdalin at doses of 0.6 and 3 mg/kg b.w. daily for 28 days. The groups S1 and S2 received commercial feed for rabbits mixed with crushed bitter apricot seeds at doses of 60 and 300 mg/kg b.w. during 28 days. The control (C) group did not receive any amygdalin. Intramuscular and peroral amygdalin administration did not affect total body weight, femoral length and femoral weight of rabbits. Similarly, microcomputed tomography (3D analysis) has shown that amygdalin had insignificant effect on relative bone volume, bone mineral density, cortical bone thickness, bone surface, trabecular thickness, trabecular number, trabecular separation. However, histological (2D analysis) revealed evident changes in compact bone microstructure of amygdalin-exposed rabbits consistent with a different vascularization and changed biomechanical properties. We can conclude that subacute exposure to amygdalin (both intramuscular and peroral) at the doses used in our study influenced compact bone remodeling

Cornelian Cherry Pulp Has Beneficial Impact on Dyslipidemia and Reduced Bone Quality in Zucker Diabetic Fatty Rats

Cornelian cherry (Cornus mas L.) is a medicinal plant with a range of biological features. It is often used as a nutritional supplement in the treatment of diabetes mellitus. Our study was aimed to first investigate the effects of Cornelian cherry pulp on bone quality parameters in Zucker diabetic fatty (ZDF) rats. Moreover, lipid-lowering properties of this fruit were also evaluated. Adult rats (n = 28) were assigned into four groups of seven individuals each: L group (non-diabetic lean rats), C group (diabetic obese rats), and E1 and E2 groups (diabetic obese rats receiving 500 and 1000 mg/kg body weight of Cornelian cherry pulp, respectively, for 10 weeks). Significantly lower levels of triglyceride, total cholesterol and alkaline phosphatase activity were determined in the E2 group versus the C group. A higher dose of Cornus mas also had a beneficial impact on femoral weight, cortical bone thickness, relative volume of trabecular bone and trabecular thickness. We observed elevated density of Haversian systems and accelerated periosteal bone apposition in both treated groups (E1 and E2). Our results clearly demonstrate that Cornelian cherry pulp has a favorable effect on lipid disorder and impaired bone quality consistent with type 2 diabetes mellitus in a suitable animal model