scholarly journals Nipah Virus Envelope-Pseudotyped Lentiviruses Efficiently Target ephrinB2-Positive Stem Cell Populations In Vitro and Bypass the Liver Sink When Administered In Vivo

2012 ◽  
Vol 87 (4) ◽  
pp. 2094-2108 ◽  
Author(s):  
K. Palomares ◽  
F. Vigant ◽  
B. Van Handel ◽  
O. Pernet ◽  
K. Chikere ◽  
...  
Keyword(s):  
Stem Cell ◽  
Nipah Virus ◽  
Cell Populations ◽  
Virus Envelope ◽  
Stem Cell Populations

scholarly journals Nipah Virus Envelope-Pseudotyped Lentiviruses Efficiently Target ephrinB2-Positive Stem Cell Populations In Vitro and Bypass the Liver Sink When Administered In Vivo

2013 ◽  
Vol 87 (8) ◽  
pp. 4794-4794
Author(s):  
K. Palomares ◽  
F. Vigant ◽  
B. Van Handel ◽  
O. Pernet ◽  
K. Chikere ◽  
...  
Keyword(s):  
Stem Cell ◽  
Nipah Virus ◽  
Cell Populations ◽  
Virus Envelope ◽  
Stem Cell Populations

Leukemia Stem Cell Potential of Different Progenitor Subpopulations in Myeloid Blast Phase CML

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3489-3489
Author(s):  
Ross Kinstrie ◽  
Dimitris Karamitros ◽  
Nicolas Goardon ◽  
Heather Morrison ◽  
Richard E Clark ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Research Funding ◽  
Cell Populations ◽  
Novel Therapies ◽  
Blast Phase ◽  
Self Renewal ◽  
Stem Cell Populations

Abstract Blast phase (BP)-CML remains the most critical area of unmet clinical need in the management of CML and novel, targeted therapeutic strategies are urgently needed. In the tyrosine kinase inhibitor (TKI) era, the rate of progression to BP is 1 to 1.5% per annum in the first few years after diagnosis, falling sharply when major molecular response is obtained. Around 10% of patients present with de novo BP-CML and despite the use of TKIs, median survival after the diagnosis of BP-CML is between 6.5 and 11 months.Therefore, improved understanding of the biology of BP-CML and novel therapies to prolong therapeutic responses are urgently sought. Studies of myeloid malignancies show that acquisition of tumor-associated mutations occurs principally in a step-wise manner. Initiating mutations usually originate in an hematopoietic stem cell (HSC) to give rise to preleukemic stem cell populations that expand through clonal advantage. Further mutation acquisition and/or epigenetic changes then lead to blast transformation and disruption of the normal immunophenotypic and functional hematopoietic hierarchy. At this stage, multiple leukemic stem cell (LSC) populations (also termed leukemia initiating cell populations) can be identified. We previously showed, in AML, that the CD34+ LSC populations were most closely related to normal progenitor populations, rather than stem cell populations, but had co-opted elements of a normal stem cell expression signature to acquire abnormal self-renewal potential (Goardon et al, Cancer Cell, 2011). CD34+CD38- LSCs were most commonly similar to an early multi-potent progenitor population with lympho-myeloid potential (the lymphoid-primed multi-potential progenitor [LMPP]). In contrast, the CD34+CD38+ LSCs were most closely related to the more restricted granulocyte-macrophage progenitor (GMP). In chronic phase CML, the leukemia-propagating population is the HSC, and the progenitor subpopulations do not have stem cell characteristics. To date, studies to isolate LSC populations in BP-CML have been limited, identifying the GMP subpopulation only as a possible LSC source (Jamieson et al, NEJM, 2004). Furthermore, in vivo LSC activity has not been assessed. We therefore set out to assess the LSC characteristics of different primitive progenitor subpopulations in myeloid BP-CML both in vitro and in vivo. We isolated different stem and progenitor cell subpopulations using FACS; HSC (Lin-CD34+CD38-CD90+ CD45RA-), multipotent progenitor (MPP; Lin-CD34+CD38-CD90-CD45RA-), LMPP (Lin-CD34+CD38-CD90-CD45RA+), common myeloid progenitor (CMP; Lin-CD34+CD38+CD45RA-CD123+), GMP (Lin-CD34+CD38+CD45RA+CD123+) and megakaryocyte erythroid progenitor (MEP; Lin-CD34+CD38+CD45RA-CD123-). The functional potential of these purified populations was examined in 13 patients by: (i) serial CFC replating assays to study progenitor self-renewal (n=10); (ii) In vivo xenograft studies using NSG mice with serial transplantation to identify populations with LSC potential (n=6). Our data conclusively demonstrate that functional LSCs are present in multiple immunophenotypic stem/progenitor subpopulations in myeloid BP-CML, including HSC, MPP, LMPP, CMP and GMP subpopulations. There was inter-patient variability in terms of both in vitro and in vivo functional properties. Fluorescence in situ hybridisation (FISH) was used to assess clonality in the different progenitor subpopulations and identify which populations contained cells with additional cytogenetic abnormalities (ACAs) with a view to improving our understanding of the clonal hierarchy. Interestingly, there were no significant differences in ACAs in the different progenitor subpopulations in the majority of samples studied, suggesting that clonal evolution tends to occur in the HSC compartment in myeloid BP-CML. Preliminary gene expression profiling studies of the different progenitor subpopulations, using Affymetrix Human Gene 1.0 ST Arrays, demonstrated highly variable gene expression, supporting the functional heterogeneity seen. Taken together, our results demonstrate that myeloid BP-CML is a very heterogeneous disorder with variable LSC populations. Further interrogation of these populations will likely identify novel therapies which will specifically target the LSC. Disclosures Copland: Bristol-Myers Squibb: Consultancy, Honoraria, Other, Research Funding; Novartis: Consultancy, Honoraria, Other; Ariad: Consultancy, Honoraria, Research Funding.

In Vitro And in Vivo Expansion of Stem Cell Populations

Vox Sanguinis ◽  
1998 ◽  
Vol 74 (S2) ◽  
pp. 463-466
Author(s):  
G. Wagemaker
Keyword(s):  
Stem Cell ◽  
Cell Populations ◽  
Stem Cell Populations

Stem Cells in Paediatric Leukaemia Are Resistant to Parthenolide.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1429-1429
Author(s):  
Charlotte Victoria Cox ◽  
Paraskevi Diamanti ◽  
Allison Blair
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Functional Capacity ◽  
Cell Populations ◽  
Minimal Effect ◽  
Childhood All ◽  
Stem Cell Populations

Abstract Abstract 1429 Poster Board I-452 The concept of cancer stem cells as developmentally early cells that are capable of continued growth and expansion in haematopoietic malignancies and solid tumours has been substantiated in recent years. Consequently these cells may be responsible for disease maintenance and relapse. Acute lymphoblastic leukaemia (ALL) is the most common paediatric cancer with survival rates around 80-85%. However, a significant proportion of patients relapse, often with disease that is highly refractory to further therapeutic intervention. Leukaemia stem cells have been described in childhood ALL that can proliferate to initiate and sustain the disease in vivo. In addition these leukaemia stem cells have also been shown to be refractory to commonly used clinical agents. Therefore it is important to investigate ALL stem cells to understand their biological properties and to identify the most appropriate agents that are capable of eradicating these cells. The sesquiterpene lactone Parthenolide (PTL) has been shown to induce apoptosis in malignant cells by inducing oxidative stress and inhibiting NF-κB activity. Importantly PTL has been shown to be effective against stem cell populations in acute myeloid leukaemia and in chronic lymphocytic leukaemia with minimal effect on normal haemopoietic cells. In this study we have attempted to assess the effects of PTL on stem cell populations in paediatric ALL. Primary cells from 20 childhood ALL cases from mixed prognostic subgroups were used in this investigation. Cells from B-ALL cases were sorted on the basis of expression of CD34/CD19, while CD34/CD7 antigens were used to sort cells from T-ALL cases. Sorted and unsorted populations were co-cultured with and without PTL at 7.5μM and 10μM for 18-24 hours. Subsequently cell viability and apoptosis were determined by flow cytometry using Annexin V and PI staining. Antibodies against phosphorylated IKKα and IKKβ were used to assess NF-κB activity in treated and untreated cells. The functional ability of the treated cells was assessed in some cases using long-term in vitro and in vivo assays. Both concentrations of PTL resulted in a significant reduction in viability in unsorted ALL cells (28±4% and 23±5% respectively). Similar results were observed with CD34+/CD19+, CD34+/CD7+ and CD34- subfractions, with viability reduced to 14-39%. In contrast the phenotypically primitive CD34+/CD19- (85±11% viable) and CD34+/CD7- (83±5% viable) populations were significantly more resistant to 10μM PTL than unsorted cells and other sorted populations (P≤0.002). FISH analyses were performed at the end of the time-course and confirmed that leukaemia cells were surviving PTL treatment. It was not possible to detect phosphorylated IKKα/β in the CD34+/CD19- and CD34+/CD7- populations, in cases examined to date, suggesting NF-kB may not be active in these subpopulations. Of note PTL treatment seemed to have minimal effect on the long-term proliferative ability of ALL cells. There were no significant differences in the absolute cell numbers generated in cultures of PTL treated CD34+/CD19- or CD34+/CD7- cells compared to untreated cells at all time points assayed up to the end of culture at week 6 (P≥0.23). Interestingly, similar results were observed with the unsorted cells and all other sorted populations. From week 3 of culture there was no difference in the absolute cell counts when growth from treated and untreated cells was compared (P>0.47), albeit they proliferated to a much lesser extent than the phenotypically primitive populations. In addition PTL treated cells were capable of engrafting NOD/SCID mice. The levels of leukaemia engraftment obtained using PTL treated unsorted (0.2-5% CD45+), CD34+/CD19- (2-10% CD45+) and CD34+/CD7- (1.5-9% CD45+) populations were similar to their respective untreated controls. These data demonstrate that while PTL showed promising effects on the bulk leukaemia cells, the effects on CD34+/CD19- B-ALL cells and CD34+/CD7- T-ALL cells were insignificant. This may be due in part to lack of NF-kB activity in leukaemia stem cells. However, the functional capacity of every ALL population evaluated in vitro was not significantly impaired by the short course of PTL treatment. These findings further highlight the importance of evaluating new therapeutic agents on leukaemia stem cell populations in addition to the bulk leukaemia and the significance of investigating the functional capacity of drug treated cells. Disclosures No relevant conflicts of interest to declare.

Heterogeneous Human Periodontal Ligament-Committed Progenitor and Stem Cell Populations Exhibit a Unique Cementogenic Property Under In Vitro and In Vivo Conditions

2017 ◽  
Vol 26 (9) ◽  
pp. 632-645 ◽  
Author(s):  
Rei Shinagawa-Ohama ◽  
Mai Mochizuki ◽  
Yuichi Tamaki ◽  
Naoto Suda ◽  
Taka Nakahara
Keyword(s):  
Stem Cell ◽  
Periodontal Ligament ◽  
Cell Populations ◽  
Stem Cell Populations

scholarly journals Activation of two distinct Sox9-EGFP-expressing intestinal stem cell populations during crypt regeneration after irradiation

2012 ◽  
Vol 302 (10) ◽  
pp. G1111-G1132 ◽  
Author(s):  
Laurianne Van Landeghem ◽  
M. Agostina Santoro ◽  
Adrienne E. Krebs ◽  
Amanda T. Mah ◽  
Jeffrey J. Dehmer ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Populations ◽  
Intestinal Epithelial ◽  
P53 Signaling ◽  
Reporter Mice ◽  
Radiation Induced ◽  
Molecular Phenotypes ◽  
Stem Cell Populations ◽  
Induced Changes

Recent identification of intestinal epithelial stem cell (ISC) markers and development of ISC reporter mice permit visualization and isolation of regenerating ISCs after radiation to define their functional and molecular phenotypes. Previous studies in uninjured intestine of Sox9-EGFP reporter mice demonstrate that ISCs express low levels of Sox9-EGFP (Sox9-EGFP Low), whereas enteroendocrine cells (EEC) express high levels of Sox9-EGFP (Sox9-EGFP High). We hypothesized that Sox9-EGFP Low ISCs would expand after radiation, exhibit enhanced proliferative capacities, and adopt a distinct gene expression profile associated with rapid proliferation. Sox9-EGFP mice were given 14 Gy abdominal radiation and studied between days 3 and 9 postradiation. Radiation-induced changes in number, growth, and transcriptome of the different Sox9-EGFP cell populations were determined by histology, flow cytometry, in vitro culture assays, and microarray. Microarray confirmed that nonirradiated Sox9-EGFP Low cells are enriched for Lgr5 mRNA and mRNAs enriched in Lgr5-ISCs and identified additional putative ISC markers. Sox9-EGFP High cells were enriched for EEC markers, as well as Bmi1 and Hopx, which are putative markers of quiescent ISCs. Irradiation caused complete crypt loss, followed by expansion and hyperproliferation of Sox9-EGFP Low cells. From nonirradiated intestine, only Sox9-EGFP Low cells exhibited ISC characteristics of forming organoids in culture, whereas during regeneration both Sox9-EGFP Low and High cells formed organoids. Microarray demonstrated that regenerating Sox9-EGFP High cells exhibited transcriptomic changes linked to p53-signaling and ISC-like functions including DNA repair and reduced oxidative metabolism. These findings support a model in which Sox9-EGFP Low cells represent active ISCs, Sox9-EGFP High cells contain radiation-activatable cells with ISC characteristics, and both participate in crypt regeneration.

scholarly journals Potency and Fate Specification in CNS Stem Cell Populations In Vitro

2008 ◽  
Vol 3 (6) ◽  
pp. 670-680 ◽  
Author(s):  
Rea Ravin ◽  
Daniel J. Hoeppner ◽  
David M. Munno ◽  
Liran Carmel ◽  
Jim Sullivan ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Populations ◽  
Fate Specification ◽  
Stem Cell Populations

scholarly journals Adaptive Mitosis Detection in Large in vitro Stem Cell Populations using Timelapse Microscopy

2011 ◽  
pp. 49-53 ◽  
Author(s):  
Tim Becker ◽  
Daniel H. Rapoport ◽  
Amir Madany Mamlouk
Keyword(s):  
Stem Cell ◽  
Cell Populations ◽  
Mitosis Detection ◽  
Stem Cell Populations

scholarly journals Chronic AMPK inactivation slows SHH medulloblastoma progression by inhibiting mTORC1 signaling and depleting tumor stem cell populations

2021 ◽  
Author(s):  
Taylor Dismuke ◽  
Daniel S Malawsky ◽  
Hedi Liu ◽  
Jay Brenman ◽  
Andrey Tikunov ◽  
...  
Keyword(s):  
Stem Cell ◽  
Genetic Model ◽  
Cellular Heterogeneity ◽  
Tumor Stem Cell ◽  
Cell Populations ◽  
Therapeutic Effects ◽  
Conditional Expression ◽  
Mtorc1 Signaling ◽  
Stem Cell Populations

We show that inactivating AMPK in vivo in a genetic model of medulloblastoma depletes tumor stem cell populations and slows tumor progression. Medulloblastoma, the most common malignant pediatric brain tumor, grows as heterogenous communities comprising diverse types of tumor and stromal cells. We have previously shown that different types of cells in medulloblastomas show different sensitivities to specific targeted therapies. To determine if specific populations depend on AMPK, we analyzed mice with AMPK-inactivated medulloblastomas. We engineered mice with brain-wide, conditional deletion of the AMPK catalytic subunits Prkaa1 and Prkaa2 and conditional expression SmoM2, an oncogenic Smo allele that hyperactivates Sonic Hedgehog (SHH) signaling. We compared the medulloblastomas that formed in these mice to tumors that form in AMPK-intact mice with conditional SmoM2 expression. AMPK-inactivated tumors progressed more slowly, allowing longer event-free survival. AMPK inactivation altered the cellular heterogeneity, determined by scRNA-seq, increasing differentiation, decreasing tumor stem cell populations and reducing glio-neuronal multipotency. Mechanistically, AMPK inactivation altered glycolytic gene expression and decreased mTORC1 pathway activation. Hk2-deletion reproduced key aspects of the AMPK-inactivation phenotype, implicating altered glycolysis in the tumor suppressive effect of AMPK inactivation. Our results show that AMPK inactivation impairs tumor growth through mechanisms that disproportionately affect tumor stem cell populations. As stem cells are intrinsically resistant to current cytotoxic therapy that drives recurrence, finding ways to target these populations may prevent treatment failure. Our data suggest that targeted AMPK inactivation may produce therapeutic effects in tumor stem cell populations refractory to other therapeutic approaches.

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