Abnormalities in reparative function of lung-derived mesenchymal stromal cells in emphysema

Mesenchymal stromal cells (MSCs) may provide crucial support in the regeneration of destructed alveolar tissue (emphysema) in COPD. We hypothesized that lung-derived MSCs (LMSCs) from emphysema patients are hampered in their repair capacity, either intrinsically or due to their interaction with the damaged micro-environment. LMSCs were isolated from lung tissue of controls and severe emphysema patients, and characterized at baseline. Additionally, LMSCs were seeded onto control and emphysematous decellularized lung tissue scaffolds and assessed for deposition of extracellular matrix (ECM). We observed no differences in surface markers, differentiation/proliferation potential and expression of ECM genes between control- and COPD-derived LMSCs. Notably, COPD-derived LMSCs displayed lower expression of FGF10 and HGF mRNA, and HGF and decorin protein. When seeded on control decellularized lung tissue scaffolds, control and COPD-derived LMSCs showed no differences in engraftment, proliferation or survival within 2 weeks, with similar ability to deposit new matrix on the scaffolds. Moreover, LMSC numbers and ability to deposit new matrix was not compromised on emphysematous scaffolds. Collectively, our data show that LMSCs from COPD patients compared to controls show less expression of FGF10 mRNA, HGF mRNA and protein and decorin protein, while other features including the mRNA expression of various ECM molecules are unaffected. Furthermore, COPD-derived LMSCs are capable of engraftment, proliferation and functioning on native lung tissue scaffolds. The damaged, emphysematous micro-environment as such does not hamper the potential of LMSCs. Thus, specific intrinsic deficiencies in growth factor production by diseased LMSCs may contribute to impaired alveolar repair in emphysema.

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Paracrine Regulation of Alveolar Epithelial Damage and Repair Responses by Human Lung-Resident Mesenchymal Stromal Cells

Cells ◽

10.3390/cells10112860 ◽

2021 ◽

Vol 10 (11) ◽

pp. 2860

Author(s):

Dennis M. L. W. Kruk ◽

Marissa Wisman ◽

Jacobien A. Noordhoek ◽

Mehmet Nizamoglu ◽

Marnix R. Jonker ◽

...

Keyword(s):

Epithelial Cells ◽

Mesenchymal Stromal Cells ◽

Lung Tissue ◽

Stromal Cells ◽

Wound Repair ◽

Lung Cells ◽

Alveolar Type ◽

Type Ii ◽

Epithelial Damage ◽

Alveolar Epithelial

COPD is characterized by irreversible lung tissue damage. We hypothesized that lung-derived mesenchymal stromal cells (LMSCs) reduce alveolar epithelial damage via paracrine processes, and may thus be suitable for cell-based strategies in COPD. We aimed to assess whether COPD-derived LMSCs display abnormalities. LMSCs were isolated from lung tissue of severe COPD patients and non-COPD controls. Effects of LMSC conditioned-medium (CM) on H2O2-induced, electric field- and scratch-injury were studied in A549 and NCI-H441 epithelial cells. In organoid models, LMSCs were co-cultured with NCI-H441 or primary lung cells. Organoid number, size and expression of alveolar type II markers were assessed. Pre-treatment with LMSC-CM significantly attenuated oxidative stress-induced necrosis and accelerated wound repair in A549. Co-culture with LMSCs supported organoid formation in NCI-H441 and primary epithelial cells, resulting in significantly larger organoids with lower type II-marker positivity in the presence of COPD-derived versus control LMSCs. Similar abnormalities developed in organoids from COPD compared to control-derived lung cells, with significantly larger organoids. Collectively, this indicates that LMSCs’ secretome attenuates alveolar epithelial injury and supports epithelial repair. Additionally, LMSCs promote generation of alveolar organoids, with abnormalities in the supportive effects of COPD-derived LMCS, reflective of impaired regenerative responses of COPD distal lung cells.

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Bone Marrow, Adipose, and Lung Tissue-Derived Murine Mesenchymal Stromal Cells Release Different Mediators and Differentially Affect Airway and Lung Parenchyma in Experimental Asthma

Stem Cells Translational Medicine ◽

10.1002/sctm.16-0398 ◽

2017 ◽

Vol 6 (6) ◽

pp. 1557-1567 ◽

Cited By ~ 34

Author(s):

Soraia C. Abreu ◽

Mariana A. Antunes ◽

Debora G. Xisto ◽

Fernanda F. Cruz ◽

Vivian C. Branco ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Lung Tissue ◽

Stromal Cells ◽

Lung Parenchyma

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Mesenchymal Stromal Cells From Myelodysplastic Syndrome Patients Show Lower DICER1 and DROSHA Expression, as Well as Decreased Mir-155 and Mir-181a MicroRNA Expression, When Compared with Healthy Controls

Blood ◽

10.1182/blood.v116.21.397.397 ◽

2010 ◽

Vol 116 (21) ◽

pp. 397-397

Author(s):

Carlos Santamaría ◽

Olga López-VIllar ◽

Sandra Muntión ◽

Belén Blanco ◽

Soraya Carrancio ◽

...

Keyword(s):

Gene Expression ◽

Bone Marrow ◽

Protein Expression ◽

Myelodysplastic Syndrome ◽

Western Blot ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Microrna Expression ◽

Healthy Controls ◽

Lower Expression

Abstract Abstract 397 Mesenchymal stromal cells (MSC) are closely related to the regulation of hematopoietic stem cell niche. Recently, Raaijmakers et al (Nature, 2010), published that deletion of Dicer1, a RNase III enzyme involved in microRNA biogenesis, in murine MSC-derived osteoprogenitors triggered peripherical blood cytopenias, myelodysplasia and subsequent AML, showing that molecular alterations in bone marrow microenvironment could result in clonal impaired haematopoiesis. Here, we have investigated whether MSC from myelodysplastic syndrome (MDS) patients show differences in DICER1 and DROSHA, another RNA III endonuclease, in comparison to healthy MSC. In addition, we have analyzed several hematopoietic-related microRNAs in these same samples. Bone marrow MSC from MDS patients (n=35; 10 5q- syndrome, 4 RA, 5 RARS, 10 RCMD, 3 RAEB, 2 MDS-U and 1 hypocellular MDS) and healthy donors (HD, n=20) were isolated and in vitro expanded following standard procedures until the third passage. Additionally, paired mononuclear cells (MNC) from 13 MDS and 8 HD were obtained. Total RNA was isolated using TRIzol reagent (Invitrogen). DICER1 and DROSHA relative gene expressions were assessed by quantitative PCR (Q-PCR) using commercial TaqMan® assay (Applied Biosystems®) with GAPDH as control gene. DICER1 and DROSHA (Abcam) protein expression were evaluated in whole cell lysates by western blot, using calnexin (Stressgen) as control. Several microRNAs with known role in hematopoiesis and immune system regulation were analyzed in 25 MDS and 12 HD by Q-PCR using commercial TaqMan® MicroRNA assay (Applied Biosystems®) with RNU43 as control microRNA. MSC from MDS showed significant lower DICER1 (0.0035±0.0020 vs. 0.0076±0.0092; p=0.044) and DROSHA (0.0070±0.0028 vs. 0.0135±0.0176; p=0.019) gene expression levels than healthy controls. Moreover, MSC from MDS showed lower protein expression of both DICER1 and DROSHA by western blot analysis, confirming Q-PCR findings. By contrast, no difference in either DICER1 (0.0197±0.0151 vs. 0.0173±0.0112; p=0.9) or DROSHA (0.0089±0.0023 vs. 0.0067±0.0037; p=0.09) gene expression were observed between MNC from MDS and HD. As far as microRNA expression, we observed a lower expression of mir-155 (0.63±0.92 vs. 0.94±0.49; p=0.007) and mir-181a (1.30±0.95 vs. 2.02±1.05; p=0.041) in MSC from MDS in comparison to healthy controls. Mir-155 and mir-181a are involved in T-cell and B-cell differentiation, while mir-155 are also related to erythroid and megakarycytic differentiation. We conclude that MSC from MDS patients show lower expression of DICER and DROSHA, two relevant RNA-III endonucleases involved in the microRNA biogenesis, confirming recent findings in murine models. Moreover, the expression of some microRNA is impaired in these cells, raising the possibility that these microenvironmental alterations could be involved in the MDS pathophysiology. Disclosures: No relevant conflicts of interest to declare.

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Lenalidomide Differentially Modifies the Genomic Profile and miRNA Expression of Mesenchymal Stromal Cells From Patients with 5q- Syndrome,

Blood ◽

10.1182/blood.v118.21.3810.3810 ◽

2011 ◽

Vol 118 (21) ◽

pp. 3810-3810

Author(s):

Sandra Muntión ◽

Carlos Santamaría ◽

Beatriz Roson ◽

Carlos Romo ◽

Olga López-Villar ◽

...

Keyword(s):

Gene Expression ◽

Gene Expression Profile ◽

Mesenchymal Stromal Cells ◽

Expression Profile ◽

Stromal Cells ◽

Microrna Expression ◽

Advisory Committees ◽

Healthy Donors ◽

Lower Expression

Abstract Abstract 3810 Mesenchymal stromal cells (MSC) are a non-hematopoietic BM cell population considered to be not only the osteoblastic progenitors, but also a key component of the hematopoietic microenvironment. Raaijmakers et al (Nature, 2010) have recently shown that deletion of Dicer1 in MSC-derived osteoprogenitors as well as its target gene SBDS resulted in myelodysplasia (MDS) in a murine model. We have previously confirmed these results in human MSC from MDS patients (ASH 2010, # 397). In a previous paper (Leukemia, 2009) we showed that MSC from 5q- syndrome patients were different from MSC from other types of MDS and could be involved in their development. We have hypothesized that lenalidomide, the standard treatment of 5q- patients could act not only on hematopoietic progenitors but also on the BM microenvironment. For this purpose BM-MSC from healthy donors (HD) (n=7) and 5q- syndrome patients (n=5) were expanded in vitro and treated with 50 uM lenalidomide or its solvent (DMSO) as control. RNA was obtained from MSC and DICER1, DROSHA and SBDS relative gene expression was assessed by real-time PCR using TaqMan® assay as well as several microRNAs with known role in hematopoiesis and immune system regulation. In addition, MSC gene expression profile was studied. Labeled samples were hybridized to affymetrix of oligonucleotide HU 1.OST arrays in 5q- patients (n=4) and compared with MSCs from HD (n=3). For this purpose the ratio lenalidomide-treated sample and its paired DMSO control was calculated and markers with a fold change >1.5 were selected for hierarchical clustering analysis (HCA). MSCs from 5q-syndrome showed lower expression of DICER1 when compared with those from HD (.35 x10−3 vs.20 x10−3 p=0.03) but this expression was recovered when 5q-MSCs were treated with Lenalidomide (0.32 x10−3 p= 0.34). By contrast, no differences in DROSHA expression were observed. In addition, 5q-MSC showed SBDS lower expression than HD-MSC and in both groups the expression increased when they were treated with lenalidomide fig1). When microRNAs were analyzed, we observed a lower microRNA expression in lenalidomide-treated MSC from healthy donors when was compared to paired non-treated cells, especially for miRNA-155 (p=0.028), miRNA-222 (p=0.028),and miRNA-181a (p=0.075; Table 1). By contrast, lenalidomide-treated MSC from MDS showed a trend towards higher microRNA expression in comparison to paired non-treated MSC.Table 1.HD-MSC DMSO vs LENA5q-MSC DMSO vs LENAmiRNA 1460.50 vs 0.30p=0.2490.07 vs 0.10p=0.7miRNA 1500.004 vs 0.0065p=0.60.001 vs 0.006p=0.07miRNA 1550.90 vs 0.58p=0.0280.80 vs 0.96p=0.7miRNA 181a2.47 vs 1,83p=0.0751.66 vs 2.32p=0.07miRNA 22286.2 vs 68.0p=0.02843.2 vs 56.2p=0.07 When the gene expression profile was carried out based in 421 selected probes including 306 known genes, MSC-treated cells from 5q- were separated from HD MSC by HCA (Fig2). We can conclude that Lenalidomide not only acts on HPC from 5q- patients but also on microenvironment by modifying the expression of DICER-1 and SBDS as well as the expression of some microRNAs and genes. Disclosures: San Miguel: Celgene Corp.: Membership on an entity's Board of Directors or advisory committees. del Cañizo:Celgene Corp.: Spanishn Adviory committee.

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Late Breaking Abstract - Aminopeptidase N/CD13 distinguishes two populations of colony-forming mesenchymal stromal cells in human lung tissue

10.1183/13993003.congress-2020.4467 ◽

2020 ◽

Author(s):

Måns Kadefors ◽

Sara Rolandsson Enes ◽

Maria Weitoft ◽

Stefan Scheding ◽

Gunilla Westergren-Thorsson

Keyword(s):

Mesenchymal Stromal Cells ◽

Lung Tissue ◽

Stromal Cells ◽

Human Lung ◽

Aminopeptidase N ◽

Human Lung Tissue ◽

Two Populations

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Mesenchymal Stromal Cells Derived from Human Non-Small Cell Lung Cancer, Normal Lung Tissue, and Bone Marrow: Differences in Frequency, Cytogenetics, Proliferation Behavior, and Chemosensitivity - Implications for Cancer Therapy.

Blood ◽

10.1182/blood.v110.11.1925.1925 ◽

2007 ◽

Vol 110 (11) ◽

pp. 1925-1925

Author(s):

Sandra Gottschling ◽

Michael Meister ◽

Anna Jauch ◽

Ulf C. Krause ◽

Anja Seckinger ◽

...

Keyword(s):

Lung Cancer ◽

Bone Marrow ◽

Small Cell Lung Cancer ◽

Mesenchymal Stromal Cells ◽

Lung Tissue ◽

Cell Lung Cancer ◽

Stromal Cells ◽

Small Cell ◽

Normal Lung ◽

Small Cell Lung

Abstract Stromal microenvironment plays a vital role for the induction and maintenance of solid tumors. Knowledge on the nature and impact of the tumor-microenvironment in lung cancer is still inadequate. In this study, we investigated and compared stromal cells (SC) derived from non-small cell lung cancer (NSCLC), normal lung tissue (NLT), and bone marrow (BM) for the presence and properties of mesenchymal stromal cells (MSC). NSCLC-SC and NLT-SC displayed mesenchymal morphology, immunophenotype, and differentiation capabilities implying their MSC character. However, their frequency, proliferation behavior, and chemosensitivity varied significantly from BM-MSC and among each other. Colony forming units-fibroblast (CFU-f) were 300-fold less frequent in bone marrow but exhibited an up to 44-fold higher colony efficiency (CE) than in lung. Compared to NLT, NSCLC specimens were four times enriched in CFU-f possessing an unfailing CE over several passages. According to colony size, the proliferation potential of NSCLC-CFU-f was double that of NLT-CFU-f and did not vary with cell density thus indicating an autonomous growth and lack of feedback regulation. Moreover, NSCLC-SC exhibited a markedly reduced sensitivity to cisplatin with delayed apoptosis and faster recovery from cytotoxic damage. In line with functional alterations multicolor-fluorescence in situ hybridization revealed genetic aberrations and chromosomal instability of NSCLC-SC but not of NLT-SC. These data provide evidence for the presence of tumor-specific MSC in NSCLC and suggest the survival of a particular tumor-stroma after chemotherapeutic treatment.

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