Young minds, deeper insights: a recap of the BMAS Summer School 2023, ranging from basic research to clinical implications of bone marrow adipose tissue.

Bone marrow adiposity (BMA) is a rapidly growing yet very young research field that is receiving worldwide attention based on its intimate relationship with skeletal and metabolic diseases, as well as hematology and cancer. Moreover, increasing numbers of young scientists and students are currently and actively working on BMA within their research projects. These developments led to the foundation of the International Bone Marrow Adiposity Society (BMAS), with the goal to promote BMA knowledge worldwide, and to train new generations of researchers interested in studying this field. Among the many initiatives supported by BMAS, there is the BMAS Summer School, inaugurated in 2021 and now at its second edition. The aim of the BMAS Summer School 2023 was to educate and train students by disseminating the latest advancement on BMA. Moreover, Summer School 2023 provided suggestions on how to write grants, deal with negative results in science, and start a laboratory, along with illustrations of alternative paths to academia. The event was animated by constructive and interactive discussions between early-career researchers and more senior scientists. In this report, we highlight key moments and lessons learned from the event.

Notch3 signaling between myeloma cells and osteocytes in the tumor niche promotes tumor growth and bone destruction.

In multiple myeloma (MM), communication via Notch signaling in the tumor niche stimulates tumor progression and bone destruction. We previously showed that osteocytes activate Notch, increase Notch3 expression, and stimulate proliferation in MM cells. We show here that Notch3 inhibition in MM cells reduced MM proliferation, decreased Rankl expression, and abrogated the ability of MM cells to promote osteoclastogenesis. Further, Notch3 inhibition in MM cells partially prevented the Notch activation and increased proliferation induced by osteocytes, demonstrating that Notch3 mediates MM-osteocyte communication. Consistently, pro-proliferative and pro-osteoclastogenic pathways were upregulated in CD138+ cells from newly diagnosed MM patients with high vs. low NOTCH3 expression. These results show that NOTCH3 signaling in MM cells stimulates proliferation and increases their osteoclastogenic potential. In contrast, Notch2 inhibition did not alter MM cell proliferation or communication with osteocytes. Lastly, mice injected with Notch3 knock-down MM cells had a 50% decrease in tumor burden and a 50% reduction in osteolytic lesions than mice bearing control MM cells. Together, these findings identify Notch3 as a mediator of cell communication among MM cells and between MM cells and osteocytes in the MM tumor niche and warrant future studies to exploit Notch3 as a therapeutic target to treat MM.

A Neuroprotective Bovine Colostrum Attenuates Apoptosis in Dexamethasone-Treated MC3T3-E1 Osteoblastic Cells.

Glucocorticoid-induced osteoporosis (GIO) is one of the most common secondary forms of osteoporosis. GIO is partially due to the apoptosis of osteoblasts and osteocytes. In addition, high doses of dexamethasone (DEX), a synthetic glucocorticoid receptor agonist, induces neurodegeneration by initiating inflammatory processes leading to neural apoptosis. Here, a neuroprotective bovine colostrum against glucocorticoid-induced neuronal damage was investigated for its anti-apoptotic activity in glucocorticoid-treated MC3T3-E1 osteoblastic cells. A model of apoptotic osteoblastic cells was developed by exposing MC3T3-E1 cells to DEX (0-700 µM). Colostrum co-treated with DEX was executed at 0.1-5.0 mg/mL. Cell viability was measured for all treatment schedules. Caspase-3 activation was assessed to determine both osteoblast apoptosis under DEX exposure and its potential prevention by colostrum co-treatment. Glutathione reduced (GSH) was measured to determine whether DEX-mediated oxidative stress-driven apoptosis is alleviated by colostrum co-treatment. Western blot was performed to determine the levels of p-ERK1/2, Bcl-XL, Bax, and Hsp70 proteins upon DEX or DEX plus colostrum exposure. Colostrum prevented the decrease in cell viability and the increase in caspase-3 activation and oxidative stress caused by DEX exposure. Cells, upon colostrum co-treated with DEX, exhibited higher levels of p-ERK1/2 and lower levels of Bcl-XL, Bax, and Hsp70. Our data support the notion that colostrum may be able to reduce DEX-induced apoptosis possibly via the activation of the ERK pathway and modulation of the Hsp70 system. We provided preliminary evidence on how bovine colostrum, as a complex and multi-component dairy product, in addition to its neuroprotective action, may affect osteoblastic cell survival undergoing apoptosis.

Targeting Notch Inhibitors to the Myeloma Bone Marrow Niche Decreases Tumor Growth and Bone Destruction without Gut Toxicity.

Systemic inhibition of Notch with γ-secretase inhibitors (GSI) decreases multiple myeloma tumor growth, but the clinical use of GSI is limited due to its severe gastrointestinal toxicity. In this study, we generated a GSI Notch inhibitor specifically directed to the bone (BT-GSI). BT-GSI administration decreased Notch target gene expression in the bone marrow, but it did not alter Notch signaling in intestinal tissue or induce gut toxicity. In mice with established human or murine multiple myeloma, treatment with BT-GSI decreased tumor burden and prevented the progression of multiple myeloma-induced osteolytic disease by inhibiting bone resorption more effectively than unconjugated GSI at equimolar doses. These findings show that BT-GSI has dual anti-myeloma and anti-resorptive properties, supporting the therapeutic approach of bone-targeted Notch inhibition for the treatment of multiple myeloma and associated bone disease. SIGNIFICANCE: Development of a bone-targeted Notch inhibitor reduces multiple myeloma growth and mitigates cancer-induced bone destruction without inducing the gastrointestinal toxicity typically associated with inhibition of Notch.

Cisplatin-Mediated Upregulation of APE2 Binding to MYH9 Provokes Mitochondrial Fragmentation and Acute Kidney Injury.

Cisplatin chemotherapy is standard care for many cancers but is toxic to the kidneys. How this toxicity occurs is uncertain. In this study, we identified apurinic/apyrimidinic endonuclease 2 (APE2) as a critical molecule upregulated in the proximal tubule cells (PTC) following cisplatin-induced nuclear DNA and mitochondrial DNA damage in cisplatin-treated C57B6J mice. The APE2 transgenic mouse phenotype recapitulated the pathophysiological features of C-AKI (acute kidney injury, AKI) in the absence of cisplatin treatment. APE2 pulldown-MS analysis revealed that APE2 binds myosin heavy-Chain 9 (MYH9) protein in mitochondria after cisplatin treatment. Human MYH9-related disorder is caused by mutations in MYH9 that eventually lead to nephritis, macrothrombocytopenia, and deafness, a constellation of symptoms similar to the toxicity profile of cisplatin. Moreover, cisplatin-induced C-AKI was attenuated in APE2-knockout mice. Taken together, these findings suggest that cisplatin promotes AKI development by upregulating APE2, which leads to subsequent MYH9 dysfunction in PTC mitochondria due to an unrelated role of APE2 in DNA damage repair. This postulated mechanism and the availability of an engineered transgenic mouse model based on the mechanism of C-AKI provides an opportunity to identify novel targets for prophylactic treatment of this serious disease. SIGNIFICANCE: These results reveal and highlight an unexpected role of APE2 via its interaction with MYH9 and suggest that APE2 has the potential to prevent acute kidney injury in patients with cisplatin-treated cancer. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/81/3/713/F1.large.jpg.

Endocrine parameters in association with bone mineral accrual in young female vocational ballet dancers.

Less is known on bone mass gains in dancers involved in vocational dance training. The present study found that, as young vocational dancers progress on their professional training, their bone health remains consistently lower compared to non-exercising controls. Endocrine mechanisms do not seem to explain these findings. PURPOSE: Little is known on bone mass development in dancers involved in vocational training. The aim of the present study was to model bone mineral content (BMC) accruals and to determine whether circulating levels of oestrogens, growth hormone (GH), and insulin-like growth factor I (IGF-1) explain differences in bone mass gains between vocational dance students and matched controls. METHODS: The total of 67 vocational female dancers (VFDs) and 68 aged-matched controls (12.1 ± 1.9 years and 12.7 ± 2.0 years at baseline, respectively) were followed for two consecutive years (34 VFD and 31 controls remained in the study for the full duration). BMC was evaluated annually at impact [femoral neck (FN); lumbar spine (LS)] and non-impact sites (forearm) using DXA. Anthropometry, age at menarche (questionnaire), and hormone serum concentrations (immunoradiometric assays) were also assessed for the same period. RESULTS: VFD demonstrated consistently reduced body weight (p < 0.001) and BMC at all three anatomical sites (p < 0.001) compared to controls throughout the study period. Menarche, body weight, GH, and IGF-1 were significantly associated with bone mass changes over time (p < 0.05) but did not explain group differences in BMC gains at impact sites (p > 0.05). However, body weight did explain the differences between groups in terms of BMC gains at the forearm (non-impact site). CONCLUSION: Two consecutive years of vocational dance training revealed that young female dancers demonstrate consistently lower bone mass compared to controls at both impact and non-impact sites. The studied endocrine parameters do not seem to explain group differences in terms of bone mass gains at impact sites.

Influence of Pilates training on muscular strength and flexibility in dancers / Influência de um programa de treino Pilates na força muscular e flexibilidade de bailarinos

The purpose of the present study was to assess the effects of a Pilates training program on muscular strength and flexibility in dance students. Fifteen dance students were divided into 2 groups: experimental (n=7) and control (n=8). Both were assessed in beginning and in the end of the study. Muscular strength was assessed measuring the time supported in the technical skills penché and developpé. To asses flexibility, it was measured the angle between limbs in the technical skills arabesque, developpé and cambré. After the first moment of evaluation, the experimental group performed a Mat-Based Pilates Exercise during 11 weeks. The statistic analyses (two-way analysis of variance - ANOVA 2x2) showed significant differences (p ≤ 0,05) in muscular strength and flexibility measurements between groups after the training program. It was concluded that Pilates training has a positive effect on muscular strength and flexibility in dance students.

Foi objectivo avaliar o efeito de um programa de treino Pilates na força muscular e flexibilidade de bailarinos estudantes. Quinze bailarinos foram divididos em 2 grupos: experimental (n=7) e controlo (n=8). Ambos foram avaliados no início e final do estudo. A força muscular foi avaliada através do tempo de sustentação nos elementos técnicos penché e developpé. Para avaliar a flexibilidade foi medido o ângulo de amplitude entre os segmentos nos elementos técnicos arabesque, cambré e developpé. Após o 1.º momento de avaliação os bailarinos do grupo experimental participaram num programa de Mat-Based Pilates Exercise durante 11 semanas. A análise estatística (análise de variância para medidas repetidas - ANOVA 2x2) demonstrou diferenças significativas (p ≤ 0,05) entre os grupos no âmbito da força muscular e flexibilidade após o programa de treino. Conclui-se que o Pilates induz alterações positivas ao nível da força muscular e flexibilidade de bailarinos.