다중혈관 관상동맥 환자에서 y-문합을 이용하여 양쪽 내흉동맥만을 사용한 우회술의 조기 성적

비만아 부모의 돌봄 경험: q 방법론

PLoS BIOLOGY Senescence-Associated Secretory Phenotypes Reveal Cell-Nonautonomous Functions of Oncogenic RAS and the p53 Tumor Suppressor Jean-Philippe Coppe 1 , Christopher K. Patil 1[ , Francis Rodier 1,2[ , Yu Sun 3 , Denise P. Mun oz 1,2 , Joshua Goldstein 1¤ , Peter S. Nelson 3 , Pierre-Yves Desprez 1,4 , Judith Campisi 1,2* 1 Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America, 2 Buck Institute for Age Research, Novato, California, United States of America, 3 Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America, 4 California Pacific Medical Center Research Institute, San Francisco, California, United States of America Cellular senescence suppresses cancer by arresting cell proliferation, essentially permanently, in response to oncogenic stimuli, including genotoxic stress. We modified the use of antibody arrays to provide a quantitative assessment of factors secreted by senescent cells. We show that human cells induced to senesce by genotoxic stress secrete myriad factors associated with inflammation and malignancy. This senescence-associated secretory phenotype (SASP) developed slowly over several days and only after DNA damage of sufficient magnitude to induce senescence. Remarkably similar SASPs developed in normal fibroblasts, normal epithelial cells, and epithelial tumor cells after genotoxic stress in culture, and in epithelial tumor cells in vivo after treatment of prostate cancer patients with DNA- damaging chemotherapy. In cultured premalignant epithelial cells, SASPs induced an epithelial–mesenchyme transition and invasiveness, hallmarks of malignancy, by a paracrine mechanism that depended largely on the SASP factors interleukin (IL)-6 and IL-8. Strikingly, two manipulations markedly amplified, and accelerated development of, the SASPs: oncogenic RAS expression, which causes genotoxic stress and senescence in normal cells, and functional loss of the p53 tumor suppressor protein. Both loss of p53 and gain of oncogenic RAS also exacerbated the promalignant paracrine activities of the SASPs. Our findings define a central feature of genotoxic stress-induced senescence. Moreover, they suggest a cell-nonautonomous mechanism by which p53 can restrain, and oncogenic RAS can promote, the development of age-related cancer by altering the tissue microenvironment. Citation: Coppe JP, Patil CK, Rodier F, Sun Y, Mun oz DP, et al. (2008) Senescence-associated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biol 6(12): e301. doi:10.1371/journal.pbio.0060301 Introduction Cancer is a multistep disease in which cells acquire increasingly malignant phenotypes. These phenotypes are acquired in part by somatic mutations, which derange normal controls over cell proliferation (growth), survival, invasion, and other processes important for malignant tumorigenesis [1]. In addition, there is increasing evidence that the tissue microenvironment is an important determinant of whether and how malignancies develop [2,3]. Normal tissue environ- ments tend to suppress malignant phenotypes, whereas abnormal tissue environments such at those caused by inﬂammation can promote cancer progression. Cancer development is restrained by a variety of tumor suppressor genes. Some of these genes permanently arrest the growth of cells at risk for neoplastic transformation, a process termed cellular senescence [4–6]. Two tumor suppressor pathways, controlled by the p53 and p16INK4a/pRB proteins, regulate senescence responses. Both pathways integrate multiple aspects of cellular physiology and direct cell fate towards survival, death, proliferation, or growth arrest, depending on the context [7,8]. Several lines of evidence indicate that cellular senescence is a potent tumor-suppressive mechanism [4,9,10]. Many poten- tially oncogenic stimuli (e.g., dysfunctional telomeres, DNA PLoS Biology | www.plosbiology.org damage, and certain oncogenes) induce senescence [6,11]. Moreover, mutations that dampen the p53 or p16INK4a/pRB pathways confer resistance to senescence and greatly increase cancer risk [12,13]. Most cancers harbor mutations in one or both of these pathways [14,15]. Lastly, in mice and humans, a senescence response to strong mitogenic signals, such as those delivered by certain oncogenes, prevents premalignant lesions from progressing to malignant cancers [16–19]. Academic Editor: Julian Downward, Cancer Research UK, United Kingdom Received June 27, 2008; Accepted October 22, 2008; Published December 2, 2008 Copyright: O 2008 Coppe et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abbreviations: CM, conditioned medium; DDR, DNA damage response; ELISA, enzyme-linked immunosorbent assay; EMT, epithelial–mesenchymal transition; GSE, genetic suppressor element; IL, interleukin; MIT, mitoxantrone; PRE, presenescent; PrEC, normal human prostate epithelial cell; REP, replicative exhaustion; SASP, senescence-associated secretory phenotype; SEN, senescent; shRNA, short hairpin RNA; XRA, X-irradiation * To whom correspondence should be addressed. E-mail: jcampisi@lbl.gov [ These authors contributed equally to this work. ¤ Current address: Genomics Institute of the Novartis Research Foundation, San Diego, California, United States of America December 2008 | Volume 6 | Issue 12 | e301

Senescence-Associated Secretory Phenotypes Reveal Cell-Nonautonomous Functions of Oncogenic RAS and the p53 Tumor Suppressor

1. Health care is a human right. 2. The care of the individual is at the center of health care, but the whole system needs to work to improve the health of populations. 3. The health care system must treat illness, alleviate suffering and disability, and promote health. 4. Cooperation with each other, those served, and those in other sectors is essential for all who work in health care. 5. All who provide health care must work to improve it. 6. Do no harm.

https://www.journalacs.org/article/S1072-7515(00)00768-7/pdf

Is this a practical approach

In recent decades, the biomedical applications of mesenchymal stem cells (MSCs) have attracted increasing attention. MSCs are easily extracted from the bone marrow, fat, and synovium, and differentiate into various cell lineages according to the requirements of specific biomedical applications. As MSCs do not express significant histocompatibility complexes and immune stimulating molecules, they are not detected by immune surveillance and do not lead to graft rejection after transplantation. These properties make them competent biomedical candidates, especially in tissue engineering. We present a brief overview of MSC extraction methods and subsequent potential for differentiation, and a comprehensive overview of their preclinical and clinical applications in regenerative medicine, and discuss future challenges.

https://www.mdpi.com/2073-4409/8/8/886/pdf

Mesenchymal Stem Cells for Regenerative Medicine.

Genipin cross-linked type II collagen/chondroitin sulfate composite hydrogel-like cell delivery system induces differentiation of adipose-derived stem cells and regenerates degenerated nucleus pulposus.

Background
Human adipose-derived mesenchymal stem cells (ADMSCs) may be ideal source of cells for intervertebral disc (IVD) regeneration, but the harsh chemical microenvironment of IVD may significantly influence the biological and metabolic vitality of ADMSCs and impair their repair potential. This study aimed to investigate the viability, proliferation and the expression of main matrix proteins of ADMSCs in the chemical microenvironment of IVD under normal and degeneration conditions.

https://link.springer.com/content/pdf/10.1186%2F1479-5876-10-49.pdf

Responses of human adipose-derived mesenchymal stem cells to chemical microenvironment of the intervertebral disc

The microenvironment of the intervertebral disc (IVD) is characterized by matrix acidity, hypoxia, hyperosmolarity and limited nutrition, which are major obstacles to stem cell-based regeneration. Our recent work showed that nucleus pulposus mesenchymal stem cells (NPMSCs) had advantages over traditional sources of cell therapy under IVD-like hypoxic and hyperosmotic conditions. Here, we examined the viability, proliferation and matrix metabolism of NPMSCs compared with adipose tissue-derived mesenchymal stem cells (ADMSCs) under IVD-like acidic conditions in vitro. ADMSCs and NPMSCs from Sprague-Dawley rats were cultured at four different pH levels representing the standard condition (pH 7.4) and the normal, mildly degenerated and severely degenerated IVD (pH 7.1, 6.8 and 6.5, respectively). Cell viability was examined by annexin-V-fluorescein isothiocyanate/propidium iodide staining. Cell proliferation was measured using a cell counting kit cell proliferation assay. The expression of aggrecan, collagen-I, collagen-II, matrix metalloproteinase-2 (MMP-2), a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS4) and the tissue inhibitor of metalloproteinase-3 (TIMP-3) was measured at mRNA and protein levels by RT-PCR and Western blotting. In both cell types, acidic pH inhibited cell viability and proliferation, downregulated the expression of aggrecan, collagen-I, collagen-II and TIMP-3, and upregulated the expression of MMP-2 and ADAMTS4. Compared with ADMSCs, NPMSCs were significantly less inhibited in viability and proliferation; they expressed significantly higher levels of aggrecan and collagen-II, and lower levels of MMP-2 and ADAMTS4. Thus, an acidic environment is a major obstacle for IVD regeneration by ADMSCs or NPMSCs. NPMSCs appeared less sensitive to inhibition by acidic pH and might be promising candidates for cell-based IVD regeneration.

Nucleus pulposus mesenchymal stem cells in acidic conditions mimicking degenerative intervertebral discs give better performance than adipose tissue-derived mesenchymal stem cells.

Adipose-derived mesenchymal stem cells (ADMSCs) and nucleus pulposus-derived mesenchymal stem cells (NPMSCs) are two cell candidates for cell-based therapies for intervertebral disc (IVD) regeneration. However, little work has been done to determine the influence of hypoxia in the IVD on the biological behaviors of ADMSCs and NPMSCs. This study aimed to investigate the viability, proliferation and differentiation of rat ADMSCs and NPMSCs in the hypoxic environment of IVD in vitro. ADMSCs and NPMSCs isolated from 6 SD rats were cultured under normoxia (20% O2) and hypoxia (2% O2) mimicking the standard condition and hypoxic environment of the IVD for 14 days. Cell viability was determined by the annexin-V-FITC/propidium iodide double-staining assay and cell proliferation was measured by MTT assay. The expression of hypoxia-inducible factor-1α, glucose transporter (GLUT)-1, GLUT-3 and vascular endothelial growth factor-A at the mRNA level was examined by RT-PCR. In cells cultured in three-dimensional micromass and differentiation medium, aggrecan, collagen-II and Sox-9 expression at mRNA and protein levels were examined by RT-PCR and Western blot. Hypoxia inhibited the viability and proliferation of both ADMSCs and NPMSCs, but promoted the chondrocytic differentiation of ADMSCs and NPMSCs. Compared to ADMSCs, NPMSCs showed greater viability, proliferation and chondrocytic differentiation under hypoxia. In conclusion, hypoxia in the IVD had a significant impact on the viability, proliferation and chondrocytic differentiation of ADMSCs and NPMSCs. NPMSCs exhibited more potent biological activity than ADMSCs in the hypoxic environment of the IVD and may represent another candidate for cell-based therapy for IVD regeneration.

https://kumc.edu/documents/msctc/li influence of hypoxia. 2013.pdf

Qixin Chen

Papers

Genipin cross-linked type II collagen/chondroitin sulfate composite hydrogel-like cell delivery system induces differentiation of adipose-derived stem cells and regenerates degenerated nucleus pulposus.

Responses of human adipose-derived mesenchymal stem cells to chemical microenvironment of the intervertebral disc

Nucleus pulposus mesenchymal stem cells in acidic conditions mimicking degenerative intervertebral discs give better performance than adipose tissue-derived mesenchymal stem cells.

Influence of Hypoxia in the Intervertebral Disc on the Biological Behaviors of Rat Adipose- and Nucleus Pulposus-Derived Mesenchymal Stem Cells

Dual release of dexamethasone and TGF-β3 from polymeric microspheres for stem cell matrix accumulation in a rat disc degeneration model.