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1.
PLoS One ; 16(2): e0246632, 2021.
Article in English | MEDLINE | ID: mdl-33591997

ABSTRACT

Adrenocortical carcinoma (ACC) is a rare but aggressive endocrine malignancy that usually results in a fatal outcome. To allow the better clinical management and reduce mortality, we searched for clinical and molecular markers that are reliable predictor of disease severity and clinical outcome in ACC patients. We determined a correlation between the overexpression of IL-13Rα2 and the clinical outcome in ACC patients using comprehensive data available in The Cancer Genome Atlas (TCGA) database. The dataset of 79 ACC subjects were divided into groups of low, medium, or high expression of IL-13Rα2 as determined by RNA-seq. These patients were also stratified by length of survival, overall survival, incidence of a new tumor event, incidence of metastasis, and production of excess hormones. We report a correlation between IL-13Rα2 expression and survival of subjects with ACC. High expression of IL-13Rα2 in ACC tumors was significantly associated with a lower patient survival rate and period of survival compared to low expression (p = 0.0084). In addition, high IL-13Rα2 expression was significantly associated with a higher incidence of new tumor events and excess hormone production compared to low or medium IL-13Rα2 expression. Within the cohort of patients that produced excess hormone, elevated IL-13Rα2 expression was significantly associated with a lower survival rate. Additionally, IL-13Rα1 had a potential relationship between transcript level and ACC survival. Our results and promising antitumor activity in preclinical models and trials indicate that IL-13Rα2 expression is an important prognostic biomarker of ACC disease outcome and a promising target for therapeutic treatment of ACC.


Subject(s)
Adrenocortical Carcinoma/genetics , Interleukin-13 Receptor alpha2 Subunit/genetics , Adrenal Cortex Neoplasms/genetics , Adrenocortical Carcinoma/immunology , Adrenocortical Carcinoma/mortality , Adult , Biomarkers, Tumor/genetics , Cohort Studies , Female , Gene Expression , Humans , Interleukin-13/genetics , Interleukin-13 Receptor alpha2 Subunit/metabolism , Male , Middle Aged , Prognosis , Severity of Illness Index , Transcriptome/genetics
2.
Stem Cells Transl Med ; 7(9): 664-675, 2018 09.
Article in English | MEDLINE | ID: mdl-30084545

ABSTRACT

Multipotent stromal cells (MSCs) are an attractive cell source for bone and cartilage tissue repair strategies. However, the functional heterogeneity of MSCs derived from different donors and manufacturing conditions has limited clinical translation, emphasizing the need for improved methods to assess MSC chondrogenic capacity. We used functionally relevant morphological profiling to dynamically monitor emergent morphological phenotypes of chondrogenically induced MSC aggregates to identify morphological features indicative of MSC chondrogenesis. Toward this goal, we characterized the morphology of chondrogenically stimulated MSC aggregates from eight different human cell-lines at multiple passages and demonstrated that MSC aggregates exhibited unique morphological dynamics that were both cell line- and passage-dependent. This variation in 3D morphology was shown to be informative of long-term MSC chondrogenesis based on multiple quantitative functional assays. We found that the specific morphological features of spheroid area, radius, minimum feret diameter, and minor axis length to be strongly correlated with MSC chondrogenic synthetic activity but not gene expression as early as day 4 in 3D culture. Our high-throughput, nondestructive approach could potentially serve as a tool to identify MSC lines with desired chondrogenic capacity toward improving manufacturing strategies for MSC-based cellular products for cartilage tissue repair. Stem Cells Translational Medicine 2018;1-12.


Subject(s)
Mesenchymal Stem Cells/metabolism , Spheroids, Cellular/metabolism , Transcriptome , Bone Marrow Cells/cytology , Cell Culture Techniques , Cell Differentiation , Cells, Cultured , Chondrogenesis , Cluster Analysis , Humans , Mesenchymal Stem Cells/cytology , Phenotype , Principal Component Analysis , Spheroids, Cellular/cytology
3.
BMC Genomics ; 18(1): 605, 2017 Aug 11.
Article in English | MEDLINE | ID: mdl-28800721

ABSTRACT

BACKGROUND: Multipotent stromal cells (MSCs) are being studied in the field of regenerative medicine for their multi-lineage differentiation and immunoregulatory capacity. MicroRNAs (miRNAs) are short non-coding RNAs that are responsible for regulating gene expression by targeting transcripts, which can impact MSC functions such as cellular proliferation, differentiation, migration and cell death. miRNAs are expressed in MSCs; however, the impact of miRNAs on cellular functions and donor variability is not well understood. Eight MSC lines were expanded to passages 3, 5 and 7, and their miRNA expression was evaluated using microarray technology. RESULTS: Statistical analyses of our data revealed that 71 miRNAs out of 939 examined were expressed by this set of MSC lines at all passages and the expression of 11 miRNAs were significantly different between passages 3 and 7, while the expression of 7 miRNAs was significantly different between passages 3 and 5. The expression of these identified miRNAs was evaluated using RT-qPCR for both the first set of MSC lines (n = 6) and a second set of MSC lines (n = 7) expanded from passages 4 to 8. By RT-qPCR only 2 miRNAs, miR-638 and miR-572 were upregulated at passage 7 compared to passage 3 in the first set of MSC lines by 1.71 and 1.54 fold, respectively; and upregulated at passage 8 compared to passage 4 in the second set of MSC lines, 1.35 and 1.59 fold, respectively. CONCLUSIONS: The expression of miR-638 and miR-572 can distinguish MSCs from two different passages of cell culture. These results may be useful in establishing critical quality attributes of MSCs and determining whether changes in these two miRNAs impact cellular functions.


Subject(s)
Gene Expression Profiling , Mesenchymal Stem Cells/metabolism , MicroRNAs/genetics , Humans , Oligonucleotide Array Sequence Analysis
4.
Stem Cells Dev ; 25(11): 861-73, 2016 06 01.
Article in English | MEDLINE | ID: mdl-27036644

ABSTRACT

Multipotent stromal cells (MSCs) are known for their distinctive ability to differentiate into different cell lineages, such as adipocytes, chondrocytes, and osteocytes. They can be isolated from numerous tissue sources, including bone marrow, adipose tissue, skeletal muscle, and others. Because of their differentiation potential and secretion of growth factors, MSCs are believed to have an inherent quality of regeneration and immune suppression. Cellular expansion is necessary to obtain sufficient numbers for use; however, MSCs exhibit a reduced capacity for proliferation and differentiation after several rounds of passaging. In this study, gene markers of MSC proliferation were identified and evaluated for their ability to predict proliferative quality. Microarray data of human bone marrow-derived MSCs were correlated with two proliferation assays. A collection of 24 genes were observed to significantly correlate with both proliferation assays (|r| >0.70) for eight MSC lines at multiple passages. These 24 identified genes were then confirmed using an additional set of MSCs from eight new donors using reverse transcription quantitative polymerase chain reaction (RT-qPCR). The proliferative potential of the second set of MSCs was measured for each donor/passage for confluency fraction, fraction of EdU+ cells, and population doubling time. The second set of MSCs exhibited a greater proliferative potential at passage 4 in comparison to passage 8, which was distinguishable by 15 genes; however, only seven of the genes (BIRC5, CCNA2, CDC20, CDK1, PBK, PLK1, and SPC25) demonstrated significant correlation with MSC proliferation regardless of passage. Our analyses revealed that correlation between gene expression and proliferation was consistently reduced with the inclusion of non-MSC cell lines; therefore, this set of seven genes may be more strongly associated with MSC proliferative quality. Our results pave the way to determine the quality of an MSC population for a particular cellular therapy in lieu of an extended in vitro or in vivo assay.


Subject(s)
Multipotent Stem Cells/cytology , Multipotent Stem Cells/metabolism , Adult , Cell Proliferation , Female , Genetic Markers , Humans , Male , Middle Aged , Oligonucleotide Array Sequence Analysis , Principal Component Analysis , Real-Time Polymerase Chain Reaction , Regression Analysis , Reproducibility of Results , Stromal Cells/cytology , Stromal Cells/metabolism , Young Adult
5.
Stem Cells ; 34(4): 935-47, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26865267

ABSTRACT

Human bone marrow-derived multipotent mesenchymal stromal cells, often referred to as mesenchymal stem cells (MSCs), represent an attractive cell source for many regenerative medicine applications due to their potential for multi-lineage differentiation, immunomodulation, and paracrine factor secretion. A major complication for current MSC-based therapies is the lack of well-defined characterization methods that can robustly predict how they will perform in a particular in vitro or in vivo setting. Significant advances have been made with identifying molecular markers of MSC quality and potency using multivariate genomic and proteomic approaches, and more recently with advanced techniques incorporating high content imaging to assess high-dimensional single cell morphological data. We sought to expand upon current methods of high dimensional morphological analysis by investigating whether short term cell and nuclear morphological profiles of MSCs from multiple donors (at multiple passages) correlated with long term mineralization upon osteogenic induction. Using the combined power of automated high content imaging followed by automated image analysis, we demonstrated that MSC morphology after 3 days was highly correlated with 35 day mineralization and comparable to other methods of MSC osteogenesis assessment (such as alkaline phosphatase activity). We then expanded on this initial morphological characterization and identified morphological features that were highly predictive of mineralization capacities (>90% accuracy) of MSCs from additional donors and different manufacturing techniques using linear discriminant analysis. Together, this work thoroughly demonstrates the predictive power of MSC morphology for mineralization capacity and motivates further studies into MSC morphology as a predictive marker for additional in vitro and in vivo responses.


Subject(s)
Bone Marrow Cells/ultrastructure , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/ultrastructure , Osteogenesis/genetics , Regenerative Medicine , Alkaline Phosphatase/metabolism , Calcification, Physiologic/genetics , Cell Differentiation/genetics , Cell Proliferation/genetics , Gene Expression Regulation, Developmental , Humans , Mesenchymal Stem Cells/metabolism , Proteomics
6.
Stem Cell Res Ther ; 5(2): 59, 2014 Apr 28.
Article in English | MEDLINE | ID: mdl-24780490

ABSTRACT

INTRODUCTION: Human multipotent stromal cells (MSCs) isolated from bone marrow or other tissue sources have great potential to treat a wide range of injuries and disorders in the field of regenerative medicine and tissue engineering. In particular, MSCs have inherent characteristics to suppress the immune system and are being studied in clinical studies to prevent graft-versus-host disease. MSCs can be expanded in vitro and have potential for differentiation into multiple cell lineages. However, the impact of cell passaging on gene expression and function of the cells has not been determined. METHODS: Commercially available human MSCs derived from bone marrow from six different donors, grown under identical culture conditions and harvested at cell passages 3, 5, and 7, were analyzed with gene-expression profiling by using microarray technology. RESULTS: The phenotype of these cells did not change as reported previously; however, a statistical analysis revealed a set of 78 significant genes that were distinguishable in expression between passages 3 and 7. None of these significant genes corresponded to the markers established by the International Society for Cellular Therapy (ISCT) for MSC identification. When the significant gene lists were analyzed through pathway analysis, these genes were involved in the top-scoring networks of cellular growth and proliferation and cellular development. A meta-analysis of the literature for significant genes revealed that the MSCs seem to be undergoing differentiation into a senescent cell type when cultured extensively. Consistent with the differences in gene expression at passage 3 and 7, MSCs exhibited a significantly greater potential for cell division at passage 3 in comparison to passage 7. CONCLUSIONS: Our results identified specific gene markers that distinguish aging MSCs grown in cell culture. Confirmatory studies are needed to correlate these molecular markers with biologic attributes that may facilitate the development of assays to test the quality of MSCs before clinical use.


Subject(s)
Mesenchymal Stem Cells/physiology , Cell Differentiation/genetics , Cell Proliferation/genetics , Cells, Cultured , Cellular Senescence/genetics , Gene Expression , Genetic Markers , Humans
7.
Am J Pathol ; 179(2): 931-41, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21684246

ABSTRACT

A wide variety of myogenic cell sources have been used for repair of injured and diseased muscle including muscle stem cells, which can be isolated from skeletal muscle as a group of slow-adhering cells on a collagen-coated surface. The therapeutic use of muscle stem cells for improving muscle regeneration is promising; however, the effect of injury on their characteristics and engraftment potential has yet to be described. In the present study, slow-adhering stem cells (SASCs) from both laceration-injured and control noninjured skeletal muscles in mice were isolated and studied. Migration and proliferation rates, multidifferentiation potentials, and differences in gene expression in both groups of cells were compared in vitro. Results demonstrated that a larger population of SASCs could be isolated from injured muscle than from control noninjured muscle. In addition, SASCs derived from injured muscle demonstrated improved migration, a higher rate of proliferation and multidifferentiation, and increased expression of Notch1, STAT3, Msx1, and MMP2. Moreover, when transplanted into dystrophic muscle in MDX/SCID mice, SASCs from injured muscle generated greater engraftments with a higher capillary density than did SASCs from control noninjured muscle. These data suggest that traumatic injury may modify stem cell characteristics through trophic factors and improve the transplantation potential of SASCs in alleviating skeletal muscle injuries and diseases.


Subject(s)
Muscle, Skeletal/metabolism , Stem Cells/cytology , Animals , Cell Adhesion , Cell Differentiation , Cell Proliferation , Flow Cytometry/methods , Gene Expression Regulation , Male , Mice , Mice, Inbred C57BL , Mice, Inbred mdx , Mice, SCID , Muscle, Skeletal/injuries , Regeneration , Satellite Cells, Skeletal Muscle/cytology , Wound Healing
8.
Int J Clin Exp Pathol ; 3(7): 681-90, 2010 Mar 25.
Article in English | MEDLINE | ID: mdl-20830239

ABSTRACT

The liver is unique for its ability to regenerate after injury, however, critical injuries or disease cause it to lose this quality. Stem cells have been explored as a possibility to restore the function of seriously damaged livers, based on their self-renewability and multiple differentiation capacity. These experiments examine the ability of muscle derived stem cells (MDSCs) to differentiate into hepatocyte-like cells in vitro and acquire functional liver attributes for repairing damaged livers. In vitro experiments were performed using MDSCs from postnatal mice and mouse hepatocyte cell lines. Our data revealed that MDSCs differentiated into hepatocyte-like cells and expressed liver cell markers, albumin, hepatocyte nuclear factor 4 α, and alpha feto-protein, both at the RNA and protein level. Additionally, in vivo studies showed successful engraftment of MDSCs into hepatectomized mouse livers of mice. These results provide evidence suggesting that MDSCs have the capacity to differentiate into liver cell-like cells and may serve as potential candidates to aid in liver regeneration.


Subject(s)
Hepatocytes/cytology , Liver Regeneration/physiology , Muscle, Skeletal/cytology , Stem Cells/cytology , Animals , Cell Differentiation/physiology , Coculture Techniques , Immunohistochemistry , In Situ Hybridization, Fluorescence , Mice , Reverse Transcriptase Polymerase Chain Reaction
9.
J Am Col Certif Wound Spec ; 2(2): 40-3, 2010 Jun.
Article in English | MEDLINE | ID: mdl-24527144

ABSTRACT

Scarring results from injuries and disease in mammalian adults and can cause pain and loss of function in the afflicted tissues. This negative aspect of wound repair is not always true for certain amphibians and during fetal development of mammals. Based on this knowledge, scientists and clinicians are investigating the mechanisms and growth factors that contribute to or deter a suitable environment for wound healing. This review summarizes these aspects and challenges for scarless repair.

10.
J Am Col Certif Wound Spec ; 1(3): 92-4, 2009 Jun 23.
Article in English | MEDLINE | ID: mdl-20161527

ABSTRACT

Three types of stem cells, embryonic, adult, and induced pluripotent stem cells, are currently studied by scientists. Barack Obama's presidency has opened the door for stem cell research by revoking statements and orders made during the former President Bush's administration. This provisional period will allow the National Institute of Health to rewrite policies governing how federal funds are distributed for stem cell research. These new regulations will grant more freedom to researchers wishing to use stem cells in their research and challenge them to determine the most appropriate stem cell treatment for a given disorder.

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