RESUMEN
The infiltration and subsequent in situ subtype specification of monocytes to effector/inflammatory and repair macrophages is indispensable for tissue repair upon acute sterile injury. However, the chromatin-level mediators and regulatory events controlling this highly dynamic macrophage phenotype switch are not known. In this study, we used a murine acute muscle injury model to assess global chromatin accessibility and gene expression dynamics in infiltrating macrophages during sterile physiological inflammation and tissue regeneration. We identified a heme-binding transcriptional repressor, BACH1, as a novel regulator of this process. Bach1 knockout mice displayed impaired muscle regeneration, altered dynamics of the macrophage phenotype transition, and transcriptional deregulation of key inflammatory and repair-related genes. We also found that BACH1 directly binds to and regulates distal regulatory elements of these genes, suggesting a novel role for BACH1 in controlling a broad spectrum of the repair response genes in macrophages upon injury. Inactivation of heme oxygenase-1 (Hmox1), one of the most stringently deregulated genes in the Bach1 knockout in macrophages, impairs muscle regeneration by changing the dynamics of the macrophage phenotype switch. Collectively, our data suggest the existence of a heme-BACH1--HMOX1 regulatory axis, that controls the phenotype and function of the infiltrating myeloid cells upon tissue damage, shaping the overall tissue repair kinetics.
Asunto(s)
Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Hemo-Oxigenasa 1/metabolismo , Proteínas de la Membrana/metabolismo , Músculo Esquelético/metabolismo , Regeneración/fisiología , Animales , Inflamación/metabolismo , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fenotipo , Transcripción Genética/fisiologíaRESUMEN
KEY POINTS: The in situ phenotypic switch of macrophages is delayed in acute injury following irradiation. The combination of bone marrow transplantation and local muscle radiation protection allows for the identification of a myeloid cell contribution to tissue repair. PET-MRI allows monitoring of myeloid cell invasion and metabolism. Altered cellular composition prior to acute sterile injury affects the in situ phenotypic transition of invading myeloid cells to repair macrophages. There is reciprocal intercellular communication between local muscle cell compartments, such as PAX7 positive cells, and recruited macrophages during skeletal muscle regeneration. ABSTRACT: Skeletal muscle regeneration is a complex interplay between various cell types including invading macrophages. Their recruitment to damaged tissues upon acute sterile injuries is necessary for clearance of necrotic debris and for coordination of tissue regeneration. This highly dynamic process is characterized by an in situ transition of infiltrating monocytes from an inflammatory (Ly6Chigh ) to a repair (Ly6Clow ) macrophage phenotype. The importance of the macrophage phenotypic shift and the cross-talk of the local muscle tissue with the infiltrating macrophages during tissue regeneration upon injury are not fully understood and their study lacks adequate methodology. Here, using an acute sterile skeletal muscle injury model combined with irradiation, bone marrow transplantation and in vivo imaging, we show that preserved muscle integrity and cell composition prior to the injury is necessary for the repair macrophage phenotypic transition and subsequently for proper and complete tissue regeneration. Importantly, by using a model of in vivo ablation of PAX7 positive cells, we show that this radiosensitive skeletal muscle progenitor pool contributes to macrophage phenotypic transition following acute sterile muscle injury. In addition, local muscle tissue radioprotection by lead shielding during irradiation preserves normal macrophage transition dynamics and subsequently muscle tissue regeneration. Taken together, our data suggest the existence of a more extensive and reciprocal cross-talk between muscle tissue compartments, including satellite cells, and infiltrating myeloid cells upon tissue damage. These interactions shape the macrophage in situ phenotypic shift, which is indispensable for normal muscle tissue repair dynamics.
Asunto(s)
Macrófagos/inmunología , Músculo Esquelético , Traumatismos Experimentales por Radiación/inmunología , Animales , Trasplante de Médula Ósea , Cardiotoxinas , Imagen por Resonancia Magnética , Masculino , Ratones Endogámicos C57BL , Músculo Esquelético/diagnóstico por imagen , Músculo Esquelético/inmunología , Músculo Esquelético/lesiones , Músculo Esquelético/efectos de la radiación , Fenotipo , Tomografía Computarizada por Tomografía de Emisión de Positrones , Traumatismos Experimentales por Radiación/diagnóstico por imagen , RegeneraciónRESUMEN
Tissue regeneration requires inflammatory and reparatory activity of macrophages. Macrophages detect and eliminate the damaged tissue and subsequently promote regeneration. This dichotomy requires the switch of effector functions of macrophages coordinated with other cell types inside the injured tissue. The gene regulatory events supporting the sensory and effector functions of macrophages involved in tissue repair are not well understood. Here we show that the lipid activated transcription factor, PPARγ, is required for proper skeletal muscle regeneration, acting in repair macrophages. PPARγ controls the expression of the transforming growth factor-ß (TGF-ß) family member, GDF3, which in turn regulates the restoration of skeletal muscle integrity by promoting muscle progenitor cell fusion. This work establishes PPARγ as a required metabolic sensor and transcriptional regulator of repair macrophages. Moreover, this work also establishes GDF3 as a secreted extrinsic effector protein acting on myoblasts and serving as an exclusively macrophage-derived regeneration factor in tissue repair.
Asunto(s)
Factor 3 de Diferenciación de Crecimiento/metabolismo , Músculo Esquelético/fisiología , Mioblastos/metabolismo , PPAR gamma/metabolismo , Regeneración/fisiología , Animales , Western Blotting , Separación Celular , Inmunoprecipitación de Cromatina , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/fisiología , Ratones , Ratones Endogámicos C57BL , Músculo Esquelético/lesiones , Análisis de Secuencia por Matrices de Oligonucleótidos , Cicatrización de Heridas/fisiologíaRESUMEN
Surgery has been considered the first choice of treatment in planocellular skin cancers. However, adjuvant radiotherapy is often required in R1 resection or in lymph node positivity. Inoperable cases are also treated with ionizing radiation with palliative purpose. The authors present a case report of a successful treatment of an 87-year-old diabetic patient with a T4N1M0 stage periauricular destructive tumour treated with 3D conformal adaptive radiotherapy. Complete remission occurred although the initial treatment aim was only palliation.
Asunto(s)
Carcinoma de Células Escamosas/radioterapia , Pabellón Auricular , Cuidados Paliativos/métodos , Radioterapia Conformacional , Neoplasias Cutáneas/radioterapia , Anciano de 80 o más Años , Carcinoma de Células Escamosas/patología , Comorbilidad , Femenino , Humanos , Estadificación de Neoplasias , Calidad de Vida , Radiodermatitis/etiología , Dosificación Radioterapéutica , Radioterapia Conformacional/efectos adversos , Radioterapia Conformacional/métodos , Inducción de Remisión , Neoplasias Cutáneas/patología , Resultado del TratamientoRESUMEN
Despite of the studies on widespread biological effects of irradiation, surprisingly only little number of papers can be found dealing with its in vivo hemorheological impact. Furthermore, other studies suggested that low-dose irradiation might differ from high-dose in more than linear ways. On Balb/c Jackson female adult mice hematological and hemorheological impacts of total body irradiation were investigated 1 hour following 0.002, 0.005, 0.01, 0.02, 0.05 and 0.1 Gy dose irradiation. In case of 0.01 Gy further groups were analyzed 30 minutes, 2, 4, 6, 24 and 48 h after irradiation. According to the results, it seems that the dose-dependent changes of blood micro-rheological parameters are not linear. The irradiation dose of 0.01 Gy acted as a point of 'inflexion', because by this dose we found the most expressed changes in hematological parameters, as well as in red blood cell aggregation, deformability and osmoscan data. The time-dependent changes showed progressive decrease in pH, rise in lactate concentration, further decrease in erythrocyte aggregation index and deformability, with moderate shifting of the optimal osmolarity point and modulation in membrane stability. As conclusion, low-dose total body irradiation may cause micro-rheological changes, being non-linearly correlated with the irradiation dose.
Asunto(s)
Agregación Eritrocitaria/efectos de la radiación , Deformación Eritrocítica/efectos de la radiación , Reología/métodos , Irradiación Corporal Total , Animales , Análisis Químico de la Sangre , Relación Dosis-Respuesta en la Radiación , Índices de Eritrocitos , Femenino , Hemorreología , Concentración de Iones de Hidrógeno , Ratones , Ratones Endogámicos BALB C , Fotones , Factores de TiempoRESUMEN
INTRODUCTION: Treatment of astrocytoma is frequently hampered by radioresistance of the tumor. In addition to overexpression of ErbB1/EGFR, functional crosstalk between receptor tyrosine kinases and cell adhesion molecules may also contribute to therapy resistance. METHODS: Acceptor photobleaching FRET was implemented on frozen sections of clinical astrocytoma to check the role of ErbB1-integrin-ß1 interaction. U251 glioma subclones were obtained by introducing extra CHR7 material or the ErbB1 gene to test the relevance and mechanism of this interaction in vitro. RESULTS: Grade IV tumors showed higher ErbB1 and integrin-ß1 expression and greater ErbB1-integrin-ß1 heteroassociation than did grade II tumors. Of these, the extent of molecular association was a single determinant of tumor grade and prognosis in stepwise logistic regression. In vitro, integrin-ß1 was upregulated, and radiosensitivity was diminished by ectopic ErbB1 expression. Great excess of ErbB1 provided colony forming advantage over medium excess but did not yield better radiation resistance or faster proliferation and decreased to medium level over time, whereas integrin-ß1 levels remained elevated and defined the extent of radioresistance. Increased expression of ErbB1 and integrin-ß1 was paralleled by decreasing ErbB1 homoassociation and increasing ErbB1-integrin-ß1 heteroassociation. Microscopic two-sided FRET revealed that pixels with higher ErbB1-integrin-ß1 heteroassociation exhibited lowed ErbB1 homoassociation, indicating competition for association partners among these molecules. Boosted Akt phosphorylation response to EGF accompanied this shift toward heteroassociation, and the consequentially increased radioresistance could be reverted by inhibiting PI3K. CONCLUSION: The clinically relevant ErbB1-integrin-ß1 heteroassociation may be used as a target of both predictive diagnostics and molecular therapy.