Cell-Surface Protein Profiling Identifies Distinctive Markers of Progenitor Cells in Human Skeletal Muscle

SummarySkeletal muscle contains two distinct stem/progenitor populations. One is the satellite cell, which acts as a muscle stem cell, and the other is the mesenchymal progenitor, which contributes to muscle pathogeneses such as fat infiltration and fibrosis. Detailed and accurate characterization of these progenitors in humans remains elusive. Here, we performed comprehensive cell-surface protein profiling of the two progenitor populations residing in human skeletal muscle and identified three previously unrecognized markers: CD82 and CD318 for satellite cells and CD201 for mesenchymal progenitors. These markers distinguish myogenic and mesenchymal progenitors, and enable efficient isolation of the two types of progenitors. Functional study revealed that CD82 ensures expansion and preservation of myogenic progenitors by suppressing excessive differentiation, and CD201 signaling favors adipogenesis of mesenchymal progenitors. Thus, cell-surface proteins identified here are not only useful markers but also functionally important molecules, and provide valuable insight into human muscle biology and diseases

Cirurgia de revascularização do miocárdio: complicações pleuro-pulmonares agudas caracterizadas por tomografia computadorizada de tórax

INTRODUCTION: Pleuropulmonary changes are common following coronary artery bypass grafting surgery performed with a saphenous vein graft, with or without an internal mammary artery. The presence of atelectasis or pleural effusions reflects the thoracic trauma. PURPOSE: To define the postoperative incidence of changes in the lung and in the pleural space and to evaluate the influence of the trauma. METHODS: Thirty patients underwent elective coronary artery bypass grafting surgery (8 saphenous vein grafts and 22 saphenous vein grafts and internal mammary artery grafts with pleurotomy). Chest tubes in the left pleural space were used in all internal mammary artery patients. On the second (day 2) and seventh (day 7) postoperative day, patients underwent a computed tomography, and pleural effusions were rated as follows: grade 0 = no fluid to grade 4 = fluid in more than 75% of the hemithorax. Atelectasis was rated as follows: laminar = 1, segmental = 3, and lobar = 10 points. RESULTS: All patients had pleural effusion or atelectasis. Between day 2 and day 7, the number of patients with effusions or atelectasis on the right side decreased (P < 0.05). The incidence of effusions on day 2 in the saphenous vein graft group (87.5%) was higher (P < 0.05) than in the internal mammary artery group (52.3%). The incidence of atelectasis in the lower right lobe decreased (P < 0.05) from 86.7% (day 2) to 26.7% (day 7). The degree of atelectasis in both sides did not differ on day 2 (P = 0.42) but did on day 7 (P < 0.0001). There was a decrease in the atelectasis from day 2 to day 7 on the right side (P < 0.001), but not on the left (P = 0.21). On day 2 there was a relationship between atelectasis and effusion on the right (P = 0.04), but not on the left (P = 0.113). CONCLUSION: The present series demonstrates that there is a high incidence of both minimal pleural effusion and atelectasis after coronary artery bypass grafting surgery, which drops on the right side from day 2 to day 7 post surgery. Factors that contribute to the persistence of changes on the left side include the thoracic trauma and the presence of chest tubes and pericardial effusion.INTRODUÇÃO: O comprometimento pleuro-pulmonar é freqüente após cirurgia de revascularização do miocárdio independente do uso de veia safena ou de artéria mamária interna. A presença de atelectasias e derrame pleural tem sido atribuída ao trauma torácico. OBJETIVOS: Definir a incidência do comprometimento pleuro-pulmonar observado através de tomografia computadorizada, analisando a influência do trauma no desenvolvimento destas complicações. MÉTODOS: Trinta pacientes foram submetidos a cirurgia eletiva de revascularização do miocárdio (8 safena e 22 também mamária com pleurotomia). Foi drenada a cavidade pleural esquerda de todos os pacientes do grupo mamária. Realizada Tomografia Computadorizada do tórax no 2º e 7º dias pós-op, avaliando-se o derrame pleural (0: sem líquido e 4: derrame em mais de 75% do hemitórax) e atelectasias (laminar: 1, segmentar: 3 e lobar: 10 pontos). RESULTADOS: Todos os pacientes apresentaram derrame pleural ou atelectasia. Entre o 2º e o 7º dia, o número de pacientes com derrame ou com atelectasia, no lado direito, diminuiu (p < 0.05). A prevalência de derrames, no 2º dia, no grupo safena, (87.5%) foi maior (p < 0.05) do que no grupo mamária (52.3%). A prevalência de atelectasias no lobo inferior direito diminuiu (p < 0.05) de 86.7% (2º dia) para 26.7% (7º dia). O grau de atelectasias, em ambos os lados, não diferiu no dia 2 (p = 0,42) mas sim no dia 7 (p < 0.0001). Houve diminuição das atelectasias do dia 2 para o 7, à direita (p < 0.001), mas não à esquerda (p = 0.21). Houve correlação (dia 2) entre as atelectasias e os derrames à direita (p = 0.04), mas não à esquerda (p = 0.113). CONCLUSÃO: Este estudo evidencia alta prevalência de derrame pleural e atelectasias após cirurgia de revascularização do miocárdio, que se reduz à direita no 7º dia de pós-operatório. Os fatores envolvidos na persis-tência das alterações à esquerda incluem o maior trauma torácico, ressaltando-se a drenagem e a presença de derrame pericárdico

Acute mediastinitis: multidetector computed tomography findings following cardiac surgery

Mediastinite pós-cirurgias torácicas é definida como a infecção dos órgãos e tecidos do espaço mediastinal, ocorrendo em 0,4% a 5% dos casos. A gravidade da infecção pós-operatória varia desde infecção de tecidos superficiais da parede torácica até mediastinite fulminante com envolvimento esternal. O critério diagnóstico da tomografia computadorizada para mediastinite aguda pós-cirúrgica é a presença de coleção mediastinal, podendo estar associada ou não a anormalidades periesternais como edema/borramento de partes moles, separação dos segmentos esternais com reabsorção óssea marginal, esclerose e osteomielite. Achados associados incluem linfonodomegalias, consolidações pulmonares e derrame pleural e pericárdico. Pequenas coleções e gás mediastinais podem ser usualmente encontradas em pós-operatório recente de cirurgias torácicas sem a presença de infecções, limitando a eficácia da tomografia computadorizada nas duas primeiras semanas. Após esse período, a tomografia alcança quase 100% de sensibilidade e especificidade. Pacientes com suspeita clínica de mediastinite devem ser submetidos a exame de tomografia para pesquisa de coleções, identificando a extensão da doença e sua natureza. A versão de multidetectores propicia recursos de reconstruções em diversos planos e janelas, contribuindo especialmente para o estudo do esterno.Postoperative mediastinitis is defined as an infection of the organs and tissues in the mediastinal space, with an incidence ranging between 0.4% and 5% of cases. This disease severity varies from infection of superficial tissues in the chest wall to fulminant mediastinitis with sternal involvement. Diagnostic criterion for postoperative detection of acute mediastinitis at computed tomography is the presence of fluid collections and gas in the mediastinal space, which might or might not be associated with peristernal abnormalities such as edema of soft tissues, separation of sternal segments with marginal bone resorption, sclerosis and osteomyelitis. Other associated findings include lymphadenomegaly, pulmonary consolidation and pleural/pericardial effusion. Some of these findings, such as mediastinal gas and small fluid collections can be typically found in the absence of infection, early in the period following thoracic surgery where the effectiveness of computed tomography is limited. After approximately two weeks, computed tomography achieves almost 100% sensitivity and specificity. Patients with clinical suspicion of mediastinitis should be submitted to computed tomography for investigating the presence of fluid collections to identify the extent and nature of the disease. Multidetector computed tomography allows 3D images reconstruction, contributing particularly to the evaluation of the sternum

Collagen-VI supplementation by cell transplantation improves muscle regeneration in Ullrich congenital muscular dystrophy model mice

6型コラーゲンを補う細胞移植がウルリッヒ型先天性筋ジストロフィーモデルマウスの病態を改善する. 京都大学プレスリリース. 2021-08-24.iPS cells show therapeutic benefits for a rare muscle dystrophy. 京都大学プレスリリース. 2021-08-24.[Background] Mesenchymal stromal cells (MSCs) function as supportive cells on skeletal muscle homeostasis through several secretory factors including type 6 collagen (COL6). Several mutations of COL6A1, 2, and 3 genes cause Ullrich congenital muscular dystrophy (UCMD). Skeletal muscle regeneration deficiency has been reported as a characteristic phenotype in muscle biopsy samples of human UCMD patients and UCMD model mice. However, little is known about the COL6-dependent mechanism for the occurrence and progression of the deficiency. The purpose of this study was to clarify the pathological mechanism of UCMD by supplementing COL6 through cell transplantation. [Methods] To test whether COL6 supplementation has a therapeutic effect for UCMD, in vivo and in vitro experiments were conducted using four types of MSCs: (1) healthy donors derived-primary MSCs (pMSCs), (2) MSCs derived from healthy donor induced pluripotent stem cell (iMSCs), (3) COL6-knockout iMSCs (COL6KO-iMSCs), and (4) UCMD patient-derived iMSCs (UCMD-iMSCs). [Results] All four MSC types could engraft for at least 12 weeks when transplanted into the tibialis anterior muscles of immunodeficient UCMD model (Col6a1KO) mice. COL6 protein was restored by the MSC transplantation if the MSCs were not COL6-deficient (types 1 and 2). Moreover, muscle regeneration and maturation in Col6a1KO mice were promoted with the transplantation of the COL6-producing MSCs only in the region supplemented with COL6. Skeletal muscle satellite cells derived from UCMD model mice (Col6a1KO-MuSCs) co-cultured with type 1 or 2 MSCs showed improved proliferation, differentiation, and maturation, whereas those co-cultured with type 3 or 4 MSCs did not. [Conclusions] These findings indicate that COL6 supplementation improves muscle regeneration and maturation in UCMD model mice

Calcitonin Receptor Signaling Inhibits Muscle Stem Cells from Escaping the Quiescent State and the Niche

Masahiko Yamaguchi, Yoko Watanabe, Takuji Ohtani, Akiyoshi Uezumi, Norihisa Mikami, Miki Nakamura, Takahiko Sato, Masahito Ikawa, Mikio Hoshino, Kunihiro Tsuchida, Yuko Miyagoe-Suzuki, Kazutake Tsujikawa, Shin’ichi Takeda, Hiroshi Yamamoto, So-ichiro Fukada, Calcitonin Receptor Signaling Inhibits Muscle Stem Cells from Escaping the Quiescent State and the Niche, Cell Reports, Volume 13, Issue 2, 2015, Pages 302-314, ISSN 2211-1247, https://doi.org/10.1016/j.celrep.2015.08.083

Cardiac side population cells have a potential to migrate and differentiate into cardiomyocytes in vitro and in vivo

Side population (SP) cells, which can be identified by their ability to exclude Hoechst 33342 dye, are one of the candidates for somatic stem cells. Although bone marrow SP cells are known to be long-term repopulating hematopoietic stem cells, there is little information about the characteristics of cardiac SP cells (CSPs). When cultured CSPs from neonatal rat hearts were treated with oxytocin or trichostatin A, some CSPs expressed cardiac-specific genes and proteins and showed spontaneous beating. When green fluorescent protein–positive CSPs were intravenously infused into adult rats, many more (∼12-fold) CSPs were migrated and homed in injured heart than in normal heart. CSPs in injured heart differentiated into cardiomyocytes, endothelial cells, or smooth muscle cells (4.4%, 6.7%, and 29% of total CSP-derived cells, respectively). These results suggest that CSPs are intrinsic cardiac stem cells and involved in the regeneration of diseased hearts

Functionally heterogeneous human satellite cells identified by single cell RNA sequencing.

Although heterogeneity is recognized within the murine satellite cell pool, a comprehensive understanding of distinct subpopulations and their functional relevance in human satellite cells is lacking. We used a combination of single cell RNA sequencing and flow cytometry to identify, distinguish, and physically separate novel subpopulations of human PAX7+ satellite cells (Hu-MuSCs) from normal muscles. We found that, although relatively homogeneous compared to activated satellite cells and committed progenitors, the Hu-MuSC pool contains clusters of transcriptionally distinct cells with consistency across human individuals. New surface marker combinations were enriched in transcriptional subclusters, including a subpopulation of Hu-MuSCs marked by CXCR4/CD29/CD56/CAV1 (CAV1+). In vitro, CAV1+ Hu-MuSCs are morphologically distinct, and characterized by resistance to activation compared to CAV1- Hu-MuSCs. In vivo, CAV1+ Hu-MuSCs demonstrated increased engraftment after transplantation. Our findings provide a comprehensive transcriptional view of normal Hu-MuSCs and describe new heterogeneity, enabling separation of functionally distinct human satellite cell subpopulations

Relayed signaling between mesenchymal progenitors and muscle stem cells ensures adaptive stem cell response to increased mechanical load

Adaptation to mechanical load, leading to enhanced force and power output, is a characteristic feature of skeletal muscle. Formation of new myonuclei required for efficient muscle hypertrophy relies on prior activation and proliferation of muscle stem cells (MuSCs). However, the mechanisms controlling MuSC expansion under conditions of increased load are not fully understood. Here we demonstrate that interstitial mesenchymal progenitors respond to mechanical load and stimulate MuSC proliferation in a surgical mouse model of increased muscle load. Mechanistically, transcriptional activation of Yes-associated protein 1 (Yap1)/transcriptional coactivator with PDZ-binding motif (Taz) in mesenchymal progenitors results in local production of thrombospondin-1 (Thbs1), which, in turn, drives MuSC proliferation through CD47 signaling. Under homeostatic conditions, however, CD47 signaling is insufficient to promote MuSC proliferation and instead depends on prior downregulation of the Calcitonin receptor. Our results suggest that relayed signaling between mesenchymal progenitors and MuSCs through a Yap1/Taz-Thbs1-CD47 pathway is critical to establish the supply of MuSCs during muscle hypertrophy