STEM-11. CD146/MCAM REGULATES MESENCHYMAL PROPERTIES, STEMNESS, RADIO-RESISTANCE AND YAP ACTIVITY IN GLIOBLASTOMA

Abstract Glioblastoma (GBM), a highly malignant and lethal brain tumor, is characterized by diffuse invasion into the brain and chemoradiotherapy resistance resulting in recurrences and poor prognosis. In this study we examined if the cell adhesion molecule CD146/ MCAM is contributing to the malignant properties of GBM, as it is known to have multiple protumorigenic functions in other tumor types. TCGA GBM database analyses revealed enhanced levels of CD146 in GBM compared to normal brain. A panel of patient-derived GBM spheroids, enriched for GBM stem cells (GSC), displayed variable and modest expression of CD146 that was strongly enhanced by cell adherence mediated by serum differentiation or ECM-coating. TGF-β-induced mesenchymal transition in U87 cells was accompanied by induction of CD146 expression, whereas ectopic overexpression of CD146/GFP in GG16 spheroids increased mesenchymal marker expression and cell invasion. Conversely, CRISPR/Cas9-generated CD146 knockouts in GSC23 spheroids led to reduced mesenchymal marker expression and invasion. These results were confirmed by applying embryonic stem cell-derived brain organoids as models to study invasion. Hence, GBM spheroid - brain organoid fusions demonstrated a stimulatory function of CD146 on invasion. Moreover, stem cell marker expression and clonogenic assays showed that CD146 increases the stem cell potential of GBM cells. The CD146 ectopic overexpression and knockout models also demonstrated involvement of CD146 in resistance to radiation that could be mechanistically linked with CD146-dependent suppression of p53 accumulation and activation of NF-kB. Interestingly, exploration of additional mechanisms potentially involved in CD146 functioning, led to the discovery that the oncogenic protein Yes-associated protein (YAP) is also regulated by CD146. Together, our findings demonstrate the importance of CD146 in controlling tumor aggressiveness and radioresistance in GBM cells.

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Taking embryonic stem cell potential to the bank: vitrification of the ICM

Journal of Assisted Reproduction and Genetics ◽

10.1007/s10815-011-9545-5 ◽

2011 ◽

Vol 28 (2) ◽

pp. 91-92

Author(s):

David F. Albertini

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Embryonic Stem ◽

Cell Potential ◽

Stem Cell Potential

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Diallyl trisulfide inhibited tobacco smoke-mediated bladder EMT and cancer stem cell marker expression via the NF-κB pathway in vivo

Journal of International Medical Research ◽

10.1177/0300060521992900 ◽

2021 ◽

Vol 49 (3) ◽

pp. 030006052199290

Author(s):

Hao Geng ◽

Wenhao Guo ◽

Lei Feng ◽

Dongdong Xie ◽

Liangkuan Bi ◽

...

Keyword(s):

Stem Cell ◽

Cancer Stem Cell ◽

Tobacco Smoke ◽

Epithelial Mesenchymal Transition ◽

Tobacco Smoke Exposure ◽

Stem Cell Marker ◽

Diallyl Trisulfide ◽

Mesenchymal Transition ◽

Marker Expression

Objective This study examined the effect of the NF-κB pathway on tobacco smoke-elicited bladder epithelial–mesenchymal transition (EMT) and cancer stem cell (CSC) marker expression in vivo. The effect of diallyl trisulfide (DATS) treatment was also examined. Methods BALB/c mice were exposed to tobacco smoke and treated with an NF-κB inhibitor and DATS. Western blotting, quantitative real-time PCR, and immunohistochemical staining were used to detect the changes of relevant indices. Results Phosphorylated inhibitor of kappa-B kinase alpha/beta expression and p65 and p50 nuclear transcription were increased by tobacco smoke exposure, whereas inhibitor of kappa-B expression was decreased. In addition, tobacco smoke reduced the expression of epithelial markers but increased that of mesenchymal and CSC markers. Our study further demonstrated that tobacco smoke-mediated EMT and CSC marker expression were attenuated by inhibition of the NF-κB pathway. Moreover, DATS reversed tobacco smoke-induced NF-κB pathway activation, EMT, and the acquisition of CSC properties in bladder tissues. Conclusions These data suggested that the NF-κB pathway regulated tobacco smoke-induced bladder EMT, CSC marker expression, and the protective effects of DATS.

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Expression of NANOG, but not POU5F1, points to the stem cell potential of primitive germ cells in neonatal pig testis

Reproduction ◽

10.1530/rep-07-0476 ◽

2008 ◽

Vol 135 (6) ◽

pp. 785-795 ◽

Cited By ~ 45

Author(s):

Sandeep Goel ◽

Mayako Fujihara ◽

Naojiro Minami ◽

Masayasu Yamada ◽

Hiroshi Imai

Keyword(s):

Stem Cell ◽

Germ Cells ◽

Embryonic Stem ◽

Large Animal ◽

Germline Development ◽

Cell Potential ◽

Transplantation Assay ◽

Neonatal Pig ◽

Stem Cell Potential ◽

Neonatal Testis

Gonocytes are primitive germ cells that are present in the neonatal testis and are committed to male germline development. Gonocytes differentiate to spermatogonia, which establish and maintain spermatogenesis in the postnatal testis. However, it is unknown whether large animal species have pluripotency-specific proteins in the testis. Nanog and Pou5f1 (Oct3/4) have been identified as transcription factors essential for maintaining pluripotency of embryonic stem cells in mice. Here, we show that NANOG protein was expressed in the germ cells of neonatal pig testes, but was progressively lost with age. NANOG was expressed in most of the lectin Dolichos biflorus agglutinin- and ZBTB16-positive gonocytes, which are known gonocyte-specific markers in pigs. NANOG was also expressed in Sertoli and interstitial cells of neonatal testes. Interestingly, POU5F1 expression was not detected at either the transcript or the protein level in neonatal pig testis. In the prepubertal testis, NANOG and POU5F1 proteins were primarily detected in differentiated germ cells, such as spermatocytes and spermatids, and rarely in undifferentiated spermatogonia. By using a testis transplantation assay, we found that germ cells from 2- to 4-day-old pigs could colonize and proliferate in the testes of the recipient mice, suggesting that primitive germ cells from neonatal pig testes have stem cell potential.

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Hematopoietic Stem Cell Purification Leads to Loss of a Stem Cell Population within the Lineage Positive Cellular Fraction

Blood ◽

10.1182/blood.v126.23.4756.4756 ◽

2015 ◽

Vol 126 (23) ◽

pp. 4756-4756

Author(s):

Laura R. Goldberg ◽

Mark Dooner ◽

Elaine Papa ◽

Mandy Pereira ◽

Del Tatto Michael ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Stem Cell ◽

Peripheral Blood ◽

Stem Cell Marker ◽

Stem Cell Population ◽

Cell Potential ◽

Hematopoietic Stem ◽

Stem Cell Potential

Abstract Background: Hematopoietic stem cells (HSCs) have tremendous self-renewal and differentiation capacity. The majority of murine hematopoietic stem cell studies have focused on rare purified populations of HSCs, conventionally described as negative for lineage-specific markers and positive for particular cell surface epitope profiles, including c-Kit, Sca-1, and CD150. However, our data indicate that such purifications lead to the loss of a significant population of actively cycling marrow cells with long-term multi-lineage stem cell potential. In the studies presented here, we tested the hypothesis that this discarded stem cell population lies, in part, within the lineage positive (Lin+) fraction of marrow. Methods: We flushed whole bone marrow (WBM) from B6.SJL mice and incubated it with allophycocyanin-tagged antibodies against erythroid (TER119), myeloid (CD11b, GR1), B-lymphoid (B220), or T-lymphoid (CD3, CD4, CD8) markers. Different doses of each specific Lin+ subset isolated by fluorescence activated cell sorting were competitively engrafted into lethally irradiated C57BL/6 host mice. At 1,3, and 6 months post-transplant, peripheral blood was analyzed for donor contribution to chimerism and lineage specificity. Results: Although typically considered to be without stem cell activity, we found that all Lin+ sub-fractions upon single sorting were able to contribute to marrow repopulation in competitive bone marrow transplants. For example, when lethally irradiated recipient mice received 3x105 C57BL/6J competitive whole bone marrow cells in combination with single-sorted GR1+ ± CD11b+ cells (2x106 cells/mouse), peripheral blood showed 15% donor chimerism at 6 months. Similarly, if single sorted CD3+ ±CD4+ ±CD8+ cells (70,000 cells/mouse), B220+ cells (1x106 cells/mouse), or Ter119+ cells (1x106 cells/mouse) were competitively engrafted with 3x105 C57BL/6 WBM cells, the donor Lin+ sub-fractions contributed to 2%, 15%, and 35% peripheral blood chimerism at 6 months post-transplant, respectively. This contribution was multi-lineage in all cases. When we performed double sorting of the Lin+ subsets, there was a dramatically reduced engraftment capacity between 1-6% donor chimerism for all subgroups. However, we do not think the loss of stem cell capacity with double sorting seen in these studies is due merely to the loss of classical hematopoietic stem cells (Lineage-/stem cell marker+). In our earlier studies, we showed that the total Lin+ population contains long-term multi-lineage engraftment capacity due almost entirely to actively cycling cells. Therefore, if the engraftment capacity within the single sorted Lin+ sub-fractions was due solely to the presence of classical HSCs lost with double sorting, the engraftment capacity found within the Lin+ compartment should be due only to quiescent cells in keeping with the cell cycle status of engrafting highly purified stem cells. Conclusions: Based on these data, we predict that a cycling population of stem cells exists within this single sorted, Lin+ enriched fraction discarded with conventional HSC purification. Future studies are ongoing to further characterize the subsets of Lin+ cells that both remain Lin+ and are found to be Lin- upon double sorting. We will analyze these populations for engraftment capacity, concomitant stem cell marker expression and cell cycle status, in order to fully characterize the total stem cell potential within whole bone marrow that is not included in the purified HSC populations. Disclosures No relevant conflicts of interest to declare.

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Distinct molecular profile and restricted stem cell potential defines the prospective human cranial neural crest from embryonic stem cell state

Stem Cell Research ◽

10.1016/j.scr.2020.102086 ◽

2020 ◽

Vol 49 ◽

pp. 102086

Author(s):

Maneeshi S. Prasad ◽

Rebekah M. Charney ◽

Lipsa J. Patel ◽

Martín I. García-Castro

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Neural Crest ◽

Embryonic Stem ◽

Molecular Profile ◽

Cranial Neural Crest ◽

Cell Potential ◽

Cell State ◽

Stem Cell Potential ◽

Stem Cell State

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Vitrification of mouse embryo derived ICM cells: a tool for preserving embryonic stem cell potential?

Fertility and Sterility ◽

10.1016/j.fertnstert.2010.07.930 ◽

2010 ◽

Vol 94 (4) ◽

pp. S240

Author(s):

N. Desai ◽

J. Xu ◽

T. Tsulaia ◽

J. Szeptycki ◽

T. Falcone ◽

...

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Mouse Embryo ◽

Embryonic Stem ◽

Cell Potential ◽

Stem Cell Potential

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Vitrification of mouse embryo-derived ICM cells: a tool for preserving embryonic stem cell potential?

Journal of Assisted Reproduction and Genetics ◽

10.1007/s10815-010-9500-x ◽

2010 ◽

Vol 28 (2) ◽

pp. 93-99 ◽

Cited By ~ 5

Author(s):

Nina Desai ◽

Jing Xu ◽

Tamara Tsulaia ◽

Julia Szeptycki-Lawson ◽

Faten AbdelHafez ◽

...

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Mouse Embryo ◽

Embryonic Stem ◽

Cell Potential ◽

Stem Cell Potential

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Canine Mesenchymal Stem Cell Potential and the Importance of Dog Breed: Implication for Cell-Based Therapies

Cell Transplantation ◽

10.3727/096368914x685294 ◽

2015 ◽

Vol 24 (10) ◽

pp. 1969-1980 ◽

Cited By ~ 18

Author(s):

Alessandro Bertolo ◽

Frank Steffen ◽

Cherry Malonzo-Marty ◽

Jivko Stoyanov

Keyword(s):

Stem Cell ◽

Mesenchymal Stem Cell ◽

Cell Potential ◽

Dog Breed ◽

Stem Cell Potential

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Differential SOD2 and GSTZ1 profiles contribute to contrasting dental pulp stem cell susceptibilities to oxidative damage and premature senescence

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02209-9 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Nadia Y. A. Alaidaroos ◽

Amr Alraies ◽

Rachel J. Waddington ◽

Alastair J. Sloan ◽

Ryan Moseley

Keyword(s):

Stem Cell ◽

Regenerative Medicine ◽

Oxidative Damage ◽

Dental Pulp ◽

Stem Cell Marker ◽

Significant Heterogeneity ◽

Premature Senescence ◽

Cell Marker ◽

Marker Expression ◽

The Impact

Abstract Background Dental pulp stem cells (DPSCs) are increasingly being advocated as viable cell sources for regenerative medicine-based therapies. However, significant heterogeneity in DPSC expansion and multi-potency capabilities are well-established, attributed to contrasting telomere profiles and susceptibilities to replicative senescence. As DPSCs possess negligible human telomerase (hTERT) expression, we examined whether intrinsic differences in the susceptibilities of DPSC sub-populations to oxidative stress-induced biomolecular damage and premature senescence further contributed to this heterogeneity, via differential enzymic antioxidant capabilities between DPSCs. Methods DPSCs were isolated from human third molars by differential fibronectin adhesion, and positive mesenchymal (CD73/CD90/CD105) and negative hematopoietic (CD45) stem cell marker expression confirmed. Isolated sub-populations were expanded in H2O2 (0–200 μM) and established as high or low proliferative DPSCs, based on population doublings (PDs) and senescence (telomere lengths, SA-β-galactosidase, p53/p16INK4a/p21waf1/hTERT) marker detection. The impact of DPSC expansion on mesenchymal, embryonic, and neural crest marker expression was assessed, as were the susceptibilities of high and low proliferative DPSCs to oxidative DNA and protein damage by immunocytochemistry. Expression profiles for superoxide dismutases (SODs), catalase, and glutathione-related antioxidants were further compared between DPSC sub-populations by qRT-PCR, Western blotting and activity assays. Results High proliferative DPSCs underwent > 80PDs in culture and resisted H2O2−induced senescence (50–76PDs). In contrast, low proliferative sub-populations exhibited accelerated senescence (4–32PDs), even in untreated controls (11-34PDs). While telomere lengths were largely unaffected, certain stem cell marker expression declined with H2O2 treatment and expansion. Elevated senescence susceptibilities in low proliferative DPSC (2–10PDs) were accompanied by increased oxidative damage, absent in high proliferative DPSCs until 45–60PDs. Increased SOD2/glutathione S-transferase ζ1 (GSTZ1) expression and SOD activities were identified in high proliferative DPSCs (10–25PDs), which declined during expansion. Low proliferative DPSCs (2–10PDs) exhibited inferior SOD, catalase and glutathione-related antioxidant expression/activities. Conclusions Significant variations exist in the susceptibilities of DPSC sub-populations to oxidative damage and premature senescence, contributed to by differential SOD2 and GSTZ1 profiles which maintain senescence-resistance/stemness properties in high proliferative DPSCs. Identification of superior antioxidant properties in high proliferative DPSCs enhances our understanding of DPSC biology and senescence, which may be exploited for selective sub-population screening/isolation from dental pulp tissues for regenerative medicine-based applications.

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