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Spatial transcriptomic interrogation of the murine bone marrow signaling landscape

Bone Research 2023年第4期11卷 847-859页

作者： Xue Xiao Conan juan Tingsheng Drennon Cedric R.Uytingco Neda Vishlaghi Dimitri Sokolowskei Lin Xu Benjamin Levi Mimi C.Sammarco robert j.tower Quantitative Biomedical Research Center Peter O’Donnell Jr.School of Public HealthUniversity of Texas Southwestern Medical CenterDallasTXUSA Department of Surgery University of Texas Southwestern Medical CenterDallasTXUSA Department of Cell Biology&Applications 10x GenomicsPleasantonCAUSA Department of Surgery Tulane School of MedicineNew OrleansLAUSA

Self-renewal and differentiation of skeletal stem and progenitor cells(SSPCs)are tightly regulated processes,with SSPC dysregulation leading to progressive bone *** the application of single-cell RNA sequencing(scRNAseq)to the bone field has led to major advancements in our understanding of SSPC heterogeneity,stem cells are tightly regulated by their neighboring cells which comprise the bone marrow ***,unbiased interrogation of these cells at the transcriptional level within their native niche environment has been ***,we combined spatial transcriptomics and scRNAseq using a predictive modeling pipeline derived from multiple deconvolution packages in adult mouse femurs to provide an endogenous,in vivo context of SSPCs within the *** combined approach localized SSPC subtypes to specific regions of the bone and identified cellular components and signaling networks utilized within the ***,the use of spatial transcriptomics allowed us to identify spatially restricted activation of metabolic and major morphogenetic signaling gradients derived from the vasculature and bone surfaces that establish microdomains within the marrow ***,we demonstrate,for the first time,the feasibility of applying spatial transcriptomics to fully mineralized tissue and present a combined spatial and single-cell transcriptomic approach to define the cellular components of the stem cell niche,identify cell-cell communication,and ultimately gain a comprehensive understanding of local and global SSPC regulatory networks within calcified tissue.

关键词： landscape utilized Spatial

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PDGFRαreporter activity identifies periosteal progenitor cells critical for bone formation and fracture repair

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Bone Research 2022年第1期10卷 122-136页

作者： jiajia Xu Yiyun Wang Zhu Li Ye Tian Zhao Li Amy Lu Ching-Yun Hsu Stefano Negri Cammy Tang robert j.tower Carol Morris Aaron W.james Departments of Pathology Johns Hopkins UniversityBaltimoreMD 21205USA Departments of Orthopedics Johns Hopkins UniversityBaltimoreMD 21205USA Department of Oral and Maxillofacial Surgery School of StomatologyChina Medical UniversityShenyangLiaoningPR China

The outer coverings of the skeleton,which is also known as the periosteum,are arranged in concentric layers and act as a reservoir for tissue-specific bone *** cellular heterogeneity within this tissue depot is being increasingly ***,inducible PDGFRαreporter animals were found to mark a population of cells within the periosteum that act as a stem cell reservoir for periosteal appositional bone formation and fracture *** these processes,PDGFRαreporter^(+)progenitors give rise to Nestin+periosteal cells before becoming osteoblasts and *** diphtheria toxin-mediated ablation of PDGFRαreporter^(+)cells led to deficits in cortical bone formation during homeostasis and a diminutive hard callus during fracture *** ossicle transplantation,both mouse PDGFRαreporter^(+)periosteal cells and human Pdgfrα+periosteal progenitors expand,ossify,and recruit marrow to a greater extent than their counterpart periosteal cells,whereas PDGFRαreporter^(−)periosteal cells exhibit a predisposition to chondrogenesis in *** RNA sequencing identified enrichment of the secreted factors Fermt3 and Ptpn6 within PDGFRαreporter^(+)periosteal cells,which partly underlie the osteoblastogenic features of this cell population.

关键词： progenitor fracture reservoir

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Neuron-to-vessel signaling is a required feature of aberrant stem cell commitment after soft tissue trauma

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Bone Research 2022年第3期10卷 615-626页

作者： Qizhi Qin Mario Gomez-Salazar Masnsen Cherief Chase A.Pagani Seungyong Lee Charles Hwang robert j.tower Sharon Onggo Yuxiao Sun Abhinav Piplani Zhao Li Sowmya Ramesh Thomas L.Clemens Benjamin Levi Aaron W.james Department of Pathology Johns Hopkins UniversityBaltimoreMD 21205USA Center for Organogenesis and Trauma Department of SurgeryUniversity of TexasSouthwesternTXUSA Department of Plastic Surgery HarvardCambridgeMA 02138USA Department of Orthopaedics Johns Hopkins UniversityBaltimoreMD 21205USA Baltimore Veterans Administration Medical Center BaltimoreMD 21201USA

The functional interdependence of nerves and blood vessels is a well-established concept during tissue morphogenesis, yet the role of neurovascular coupling in proper and aberrant tissue repair is an emerging field of interest. Here, we sought to define the regulatory relationship of peripheral nerves on vasculature in a severe extremity trauma model in mice, which results in aberrant cell fate and heterotopic ossification(HO). First, a high spatial degree of neurovascular congruency was observed to exist within extremity injury associated heterotopic ossification. Vascular and perivascular cells demonstrate characteristic responses to injury,as assessed by single cell RNA sequencing. This vascular response to injury was blunted in neurectomized mice, including a decrease in endothelial proliferation and type H vessel formation, and a downregulation of key transcriptional networks associated with angiogenesis. Independent mechanisms to chemically or genetically inhibit axonal ingrowth led to similar deficits in HO site angiogenesis, a reduction in type H vessels, and heterotopic bone formation. Finally, a combination of single cell transcriptomic approaches within the dorsal root ganglia identified key neural-derived angiogenic paracrine factors that may mediate neuron-to-vascular signaling in HO. These data provide further understanding of nerve-to-vessel crosstalk in traumatized soft tissues, which may reflect a key determinant of mesenchymal progenitor cell fate after injury.

关键词： aberrant trauma nerves

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Comparison of skeletal and soft tissue pericytes identifies CXCR4+ bone forming mural cells in human tissues

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Bone Research 2020年第3期8卷 286-299页

作者： jiajia Xu Dongqing Li Ching-Yun Hsu Ye Tian Leititia Zhang Yiyun Wang robert j.tower Leslie Chang Carolyn A.Meyers Yongxing Gao Kristen Broderick Carol Morris jody E.Hooper Sridhar Nimmagadda Bruno Peault Aaron W.james Departments of Pathology Johns Hopkins UniversityBaltimore 21205 MDUSA Department of Microbiology Wuhan University School of Basic Medical of ScienceWuhanHubeiP.R.China Department of Oral and Maxillofacial Surgery School of StomatologyChina Medical UniversityShenyangLiaoningP.R.China Departments of Orthopaedics Johns Hopkins UniversityBaltimore 21205 MDUSA Departments of Plastic Surgery Johns Hopkins UniversityBaltimore 21205 MDUSA Departments of Autopsy Service Johns Hopkins UniversityBaltimoreMD 21205USA Departments of Radiology and Radiological Science bonBaltimore 21205 MDUSA UCLA and Orthopaedic Hospital Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center Los AngelesCA 90095USA Center for Cardiovascular Science and MRC Center for Regenerative Medicine University of EdinburghEdinburghUK

Human osteogenic progenitors are not precisely defined,being primarily studied as heterogeneous multipotent cell populations and termed mesenchymal stem cells(MSCs).Notably,select human pericytes can develop into bone-forming ***,we sought to define the differentiation potential of CD146 f human pericytes from skeletal and soft tissue sources,with the underlying goal of defining cell surface markers that typify an osteoblastogenic ***146+CD31~CD45_pericytes were derived by fluorescence-activated cell sorting from human periosteum,adipose,or dermal *** CD146+CD31—CD45 cells retained canonical features of pericytes/*** pericytes demonstrated a striking tendency to undergo osteoblastogenesis in vitro and skeletogenesis in vivo,while soft tissue pericytes did not *** analysis revealed higher CXCR4 signaling among periosteal pericytes in comparison to their soft tissue counterparts,and CXCR4 chemical inhibition abrogated ectopic ossification by periosteal ***,enrichment of CXCR4+pericytes or stromal cells identified an osteoblastic/non-adipocytic precursor *** sum,human skeletal and soft tissue pericytes differ in their basal abilities to form *** exists in soft tissue pericytes,however,and CXCR4+pericytes represent an osteoblastogenic,non-adipocytic cell ***,enrichment for CXCR4-expressing stromal cells is a potential new tactic for skeletal tissue engineering.

关键词： CD146 CXCR4 pericyte

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The HIF-1α/PLOD2 axis integrates extracellular matrix organization and cell metabolism leading to aberrant musculoskeletal repair

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Bone Research 2024年第2期12卷 289-301页

作者： Heeseog Kang Amy L.Strong Yuxiao Sun Lei Guo Conan juan Alec C.Bancroft ji Hae Choi Chase A.Pagani Aysel A.Fernandes Michael Woodard juhoon Lee Sowmya Ramesh Aaron W.james David Hudson Kevin N.Dalby Lin Xu robert j.tower Benjamin Levi Center for Organogenesis Regeneration and TraumaDepartment of SurgeryUniversity of Texas SouthwesternDallasTX 75390USA Section of Plastic Surgery Department of SurgeryUniversity of MichiganAnn ArborMI 48109USA Quantitative Biomedical Research Center Peter O’Donnell Jr.School of Public HealthUniversity of Texas SouthwesternDallasTX 75390USA Department of Orthopedics and Sports Medicine University of WashingtonSeattleWA 98195USA Division of Chemical Biology and Medicinal Chemistry College of PharmacyUniversity of Texas at AustinAustinTX 78712USA Department of Pathology Johns Hopkins UniversityBaltimoreMD 21218USA

While hypoxic signaling has been shown to play a role in many cellular processes,its role in metabolism-linked extracellular matrix(ECM)organization and downstream processes of cell fate after musculoskeletal injury remains to be ***(HO)is a debilitating condition where abnormal bone formation occurs within extra-skeletal *** andhypoxia-inducible factor 1α(HIF-1α)activation have been shown to promote ***,the underlying molecular mechanisms bywhich the HIF-1αpathway in mesenchymal progenitor cells(MPCs)contributes to pathologic bone formation remain to ***,we used a proven mouse injury-induced HO model to investigate the role of HIF-1αon aberrant cell ***-cell RNA sequencing(scRNA-seq)and spatial transcriptomics analyses of the HO site,we found that collagen ECM organizationis the most highly up-regulated biological process in *** mesenchymal cell-specific deletion of Hif1α(Hoxa11-CreER^(T2);Hif1a^(fl/fl))significantly mitigated HO in ***-seq analysis of these Hoxa11-CreER^(T2);Hif1a^(fl/fl)mice identified the PLOD2/LOXpathway for collagen cross-linking as downstream of the HIF-1αregulation of ***,our scRNA-seq data and mechanisticstudies further uncovered that glucose metabolism in MPCs is most highly impacted by HIF-1α*** a translational aspect,a pan-LOX inhibitor significantly decreased HO.A newly screened compound revealed that the inhibition of PLOD2 activity in MPCssignificantly decreased osteogenic differentiation and glycolytic *** suggests that the HIF-1α/PLOD2/LOX axis linked tometabolism regulates HO-forming MPC *** results suggest that the HIF-1α/PLOD2/LOX pathway represents a promisingstrategy to mitigate HO formation.

关键词： metabolism skeletal aberrant

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Sleep as a window to understand and regulate Alzheimer's disease:emerging roles of thalamic reticular nucleus

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Neural Regeneration Research 2025年第6期20卷 1711-1712页

作者： Haoqi Sun Shiqian Shen robert j.Thomas M.Brandon Westover Can Zhang Department of Neurology Beth Israel Deaconess Medical CenterHarvard Medical SchoolBostonMAUSA Center for Translational Pain Research Department of AnesthesiaCritical Care and Pain MedicineMassachusetis General HospitalHarvard Medical SchoolBostonMAUSA Division of Pulmonary Critical Care and Sleep MedicineDepartment of MedicineBeth Israel Deaconess Medical CenterHarvard Medical SchoolBostonMAUSA Genetics and Aging Research Unit McCance Center for Brain HealthMass General Institute for Neurodegenerative DiseaseDepartment of NeurologyMassachusetts General HospitalHarvard Medical SchoolBostonMAUSA

Introduction:Alzheimer 's disease(AD) is a common neurodegenerative disorder and the primary cause of dementia. Considerable evidence supports the “amyloid hypothesis,” stating that the pathogenesis of AD is primarily caused by the deposition of amyloid-β(Aβ), which drives tau phosphorylation, neuroinflammation, and neurodegeneration in the brain. The amyloid hypothesis is strengthened by the significant and moderate benefit of lecanemab, a humanized antibody through an anti-amyloid mechanism,showing slowed clinical decline(van Dyck et al.,2023). The recent positive results of anti-amyloid trials have brought back focus on the amyloid hypothesis through biochemical, genetic, and pharmacological approaches(Zhang, 2023).

关键词： Alzheimer inflammation hypothesis

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Necrotic enteritis and antibiotic-free production of broiler chickens:Challenges in testing and using alternative products

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Animal Nutrition 2024年第1期16卷 288-298页

作者： robert j.Moore School of Science RMIT UniversityBundooraVictoria 3083Australia

The global trend towards raising broiler chickens without the use of in-feed antibiotics(IFAs)means that there is an ongoing need to develop alternative treatments capable of delivering the benefits that IFAs previously *** supported the productivity performance of chickens and played a key role in maintaining their *** enteritis(NE)is an important disease of broilers that affects health,productivity,and welfare,and was previously well controlled by ***,with the reduction in IFA use,NE is resurgent in some *** and various feed additives,including pre-,pro-,and postbiotics,phytobiotics,fatty acids,and phage therapies have been introduced as alternative methods of NE *** some of these feed additives have specific activity against the NE pathogen,Clostridium perfringens,most have the more general goal of reinforcing gut *** reviews of the effects of many of these feed additives on gut health have been published ***,rather than cover previously well reviewed areas of research this review focuses on the challenges and pitfalls in undertaking experimental assessment of alternative NE treatments and translating laboratory research to real world commercial production *** review is based on the author's particular experience,reading,thoughts,and analysis of the available information and inevitably presents a particular understanding that is likely to be at odds with others thinking on these *** is put forward to stimulate thinking and discussion on the issues covered.

关键词： Necrotic enteritis Clostridium perfringens Vaccines Feed additive Farm management

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The Orosirian(1800–2050 Ma)plate tectonic episode:Key for reconstructing the Proterozoic tectonic record

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Geoscience Frontiers 2023年第3期14卷 1-9页

作者： robert j.Stern Geosciences Dept. U Texas at DallasRichardsonTX 75083-0688United States

Eight lines of evidence indicate that the Orosirian Period in mid-Paleoproterozoic time was characterized by plate tectonics:ophiolites,low T/P metamorphism including eclogites,passive margin formation,tall mountains,paleomagnetic constraints,ore deposits,abundant S-type granites,and seismic images of paleo-subduction *** plate tectonic episode occurred about 1 billion years earlier than the present plate tectonic episode began in Neoproterozoic *** two plate tectonic episodes bracket the‘Boring Billion’,which may have been a protracted single lid tectonic episode that began when the supercontinent Nuna or Columbia *** of multiple lines of evidence for Orosirian plate tectonics demonstrates that Earth’s tectonic style can be reconstructed with some confidence back to at least Early Paleoproterozoic time,and thus the absence of compelling evidence for Mesoproterozoic plate tectonics is not obvious due to poor preservation.A tectono-magmatic lull2.3 Ga suggests an earlier episode of single lid *** for two episodes of plate tectonics and two episodes of single lid tectonics indicates that Earth switched between single lid and plate tectonics multiple times during the last 2.4 Ga.

关键词： Plate tectonics Single lid tectonics Urey ratio Paleoproterozoic

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Is plate tectonics needed to evolve technological species on exoplanets?

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Geoscience Frontiers 2016年第4期7卷 573-580页

作者： robert j.Stern Geoscience Department University of Texas at Dallas

As we continue searching for exoplanets, we wonder if life and technological species capable of communicating with us exists on any of them. As geoscientists, we can also wonder how important is the presence or absence of plate tectonics for the evolution of technological species. This essay considers this question, focusing on tectonically active roclw （silicate） planets, like Earth, Venus, and Mars. The development of technological species on Earth provides key insights for understanding evolution on exoplanets, including the likely role that plate tectonics may play. An Earth-sized silicate planet is likely to experience several tectonic styles over its lifetime, as it cools and its lithosphere thickens, strengthens, and becomes denser. These include magma ocean, various styles of stagnant lid, and perhaps plate tectonics. Abundant liquid water favors both life and plate tectonics. Ocean is required for early evolution of diverse single-celled organisms, then colonies of cells which specialized further to form guts, ap- pendages, and sensory organisms up to the complexity of fish （central nervous system, appendages, eyes）. Large expanses of dry land also begin in the ocean, today produced above subduction zones in juvenile arcs and by their coalescence to form continents, although it is not clear that plate tectonics was required to create continental crust on Earth. Dry land of continents is required for further evolution of technological species, where modification of appendages for grasping and manipulating, and improve- ment of eyes and central nervous system could be perfected. These bioassets allowed intelligent crea- tures to examine the night sky and wonder, the beginning of abstract thinking, including religion and science. Technology arises from the exigencies of daily living such as tool-making, agriculture, clothing, and weapons, but the pace of innovation accelerates once it is allied with science. Finally, the importance of plate tectonics for devel

关键词： Plate tectonics Exoplanets Planetary science Intelligent life

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Weight measurement using image-based pose analysis

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Progress in Natural Science:Materials International 2008年第12期18卷 1507-1512页

作者： robert j.Sclabassi Laboratory for Computational Neuroseience Departments of NeurosurgeryElectrical & Computer Engineeringand BioengineeringUniversity of PittsburghPittsburgh 15213USA

Image-based gait analysis as a means of biometric identification has attracted much research *** of the existing methods focus on human identification,posture analysis and movement *** have been few investigations on measuring the carried load based on the carrier’s gait characteristics by automatic image ***,this measurement is very useful in a number of applications,such as the study of the carried load on the postural development of children and *** this paper,we inves- tigate how to automatically estimate the carried weight from a sequence of *** present a method to extract human gait silhouette based on an observation that humans tend to minimize the energy during *** compute several angles of body leaning and deter- mine the relationship of the carried weight,the leaning angles and the centroid location according to a human kinetic *** weight determination method has been verified successfully by experiments.

关键词： Image-based pose analysis Energy minimization Pattern recognition Load carrying measurement

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