A dormant sub-population expressing interleukin-1 receptor characterises anti-estrogen resistant ALDH+ breast cancer stem cells

SUMMARYEstrogen receptor-positive (ER+) breast tumours are often treated with anti-estrogen (AE) therapies but frequently develop resistance. Cancer Stem Cells (CSCs) with high aldehyde dehydrogenase (ALDH) activity (ALDH+ cells) are reported to be enriched following AE treatment. Here we perform in vitro and in vivo functional CSC assays and gene expression analysis to characterise the ALDH+ population in AE resistant metastatic patient samples and an ER+ cell line. We show that the IL1β signalling pathway is activated in ALDH+ cells and data from single cells reveals that AE treatment selects for IL1R1-expressing ALDH+ cells. Importantly, we demonstrate that increased expression of IL1R1 is observed in the tumours of patients treated with AE therapy and predicts for treatment failure. Single-cell gene expression analysis revealed that at least 2 sub-populations exist within the ALDH+ population, one proliferative and one quiescent. Following AE therapy, the quiescent ALDH+IL1R1+ population is expanded, which suggests CSC dormancy as an adaptive strategy that facilitates treatment resistance. Supporting this, analysis of AE resistant dormant tumours reveals significantly increased expression of ALDH1A1, ALDH1A3 and IL1R1 genes. Thus, we propose that targeting of ALDH+IL1R1+ cells will reverse AE resistance, including in patients with minimal residual disease.

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Human and murine amniotic fluid c-Kit+Lin− cells display hematopoietic activity

Blood ◽

10.1182/blood-2008-10-182105 ◽

2009 ◽

Vol 113 (17) ◽

pp. 3953-3960 ◽

Cited By ~ 100

Author(s):

Andrea Ditadi ◽

Paolo de Coppi ◽

Olivier Picone ◽

Laetitia Gautreau ◽

Rim Smati ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Amniotic Fluid ◽

Expression Analysis ◽

Gene Expression Analysis ◽

Lymphoid Cells ◽

Immunocompromised Hosts ◽

Hematopoietic Activity

Abstract We have isolated c-Kit+Lin− cells from both human and murine amniotic fluid (AF) and investigated their hematopoietic potential. In vitro, the c-Kit+Lin− population in both species displayed a multilineage hematopoietic potential, as demonstrated by the generation of erythroid, myeloid, and lymphoid cells. In vivo, cells belonging to all 3 hematopoietic lineages were found after primary and secondary transplantation of murine c-Kit+Lin− cells into immunocompromised hosts, thus demonstrating the ability of these cells to self-renew. Gene expression analysis of c-Kit+ cells isolated from murine AF confirmed these results. The presence of cells with similar characteristics in the surrounding amnion indicates the possible origin of AF c-Kit+Lin− cells. This is the first report showing that cells isolated from the AF do have hematopoietic potential; our results support the idea that AF may be a new source of stem cells for therapeutic applications.

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100 Understanding aggressive colorectal cancers by gene expression analysis of cancer stem cells

European Journal of Cancer ◽

10.1016/s0959-8049(15)30002-2 ◽

2015 ◽

Vol 51 ◽

pp. S1

Author(s):

J. Manhas ◽

A. Bhattacharya ◽

S.K. Agrawal ◽

M. Bhat ◽

D. Ghosh ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Cancer Stem Cells ◽

Expression Analysis ◽

Gene Expression Analysis ◽

Colorectal Cancers

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15025 Correlation of in vitro gene expression analysis with in vivo efficacy

Journal of the American Academy of Dermatology ◽

10.1016/j.jaad.2020.06.646 ◽

2020 ◽

Vol 83 (6) ◽

pp. AB139

Author(s):

Tiffany Quinn ◽

Robert Harper

Keyword(s):

Gene Expression ◽

Expression Analysis ◽

Gene Expression Analysis ◽

In Vivo Efficacy

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375 Gene expression analysis of galactosamine-induced hepatotoxicity in-vivo and in-vitro

Toxicology Letters ◽

10.1016/s0378-4274(03)90374-2 ◽

2003 ◽

Vol 144 ◽

pp. s102

Author(s):

H. Hildebrand ◽

G. Kempka ◽

H. Ellinger ◽

B. Stuart ◽

B. Wahle ◽

...

Keyword(s):

Gene Expression ◽

Expression Analysis ◽

Gene Expression Analysis

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Tropism of liver epithelial cells toward hepatocellular carcinoma in vitro and in vivo with altering gene expression of cancer stem cells

The American Journal of Surgery ◽

10.1016/j.amjsurg.2017.11.041 ◽

2018 ◽

Vol 215 (4) ◽

pp. 735-743

Author(s):

Kuo-Shyang Jeng ◽

Chi-Juei Jeng ◽

Wen-Juei Jeng ◽

I-Shyan Sheen ◽

Shih-Yun Li ◽

...

Keyword(s):

Gene Expression ◽

Hepatocellular Carcinoma ◽

Stem Cells ◽

Epithelial Cells ◽

Cancer Stem Cells ◽

Liver Epithelial Cells

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Role of the snake venom toxin jararhagin in proinflammatory pathogenesis: In vitro and in vivo gene expression analysis of the effects of the toxin

Archives of Biochemistry and Biophysics ◽

10.1016/j.abb.2005.06.007 ◽

2005 ◽

Vol 441 (1) ◽

pp. 1-15 ◽

Cited By ~ 38

Author(s):

Paul Gallagher ◽

Yongde Bao ◽

Solange M.T. Serrano ◽

Gavin D. Laing ◽

R. David G. Theakston ◽

...

Keyword(s):

Gene Expression ◽

Expression Analysis ◽

Snake Venom ◽

Gene Expression Analysis ◽

Venom Toxin ◽

Snake Venom Toxin

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Gene expression analysis of human adipose tissue-derived stem cells during the initial steps of in vitro osteogenesis

Scientific Reports ◽

10.1038/s41598-018-22991-6 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 8

Author(s):

Anny Waloski Robert ◽

Addeli Bez Batti Angulski ◽

Lucia Spangenberg ◽

Patrícia Shigunov ◽

Isabela Tiemy Pereira ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Adipose Tissue ◽

Expression Analysis ◽

Gene Expression Analysis ◽

Human Adipose Tissue

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New Shuttle Vectors for Real-Time Gene Expression Analysis in Multidrug-Resistant Acinetobacter Species: In Vitro and In Vivo Responses to Environmental Stressors

Applied and Environmental Microbiology ◽

10.1128/aem.01334-19 ◽

2019 ◽

Vol 85 (18) ◽

Cited By ~ 1

Author(s):

Massimiliano Lucidi ◽

Daniela Visaggio ◽

Elisa Prencipe ◽

Francesco Imperi ◽

Giordano Rampioni ◽

...

Keyword(s):

Gene Expression ◽

Expression Analysis ◽

Gene Expression Analysis ◽

Multidrug Resistant ◽

Antibiotic Selection ◽

Content Type ◽

Shuttle Vectors ◽

Reporter Systems

ABSTRACT The Acinetobacter genus includes species of opportunistic pathogens and harmless saprophytes. The type species, Acinetobacter baumannii, is a nosocomial pathogen renowned for being multidrug resistant (MDR). Despite the clinical relevance of infections caused by MDR A. baumannii and a few other Acinetobacter spp., the regulation of their pathogenicity remains elusive due to the scarcity of adequate genetic tools, including vectors for gene expression analysis. Here, we report the generation and testing of a series of Escherichia coli-Acinetobacter promoter-probe vectors suitable for gene expression analysis in Acinetobacter spp. These vectors, named pLPV1Z, pLPV2Z, and pLPV3Z, carry both gentamicin and zeocin resistance markers and contain lux, lacZ, and green fluorescent protein (GFP) reporter systems downstream of an extended polylinker, respectively. The presence of a toxin-antitoxin gene system and the high copy number allow pLPV plasmids to be stably maintained even without antibiotic selection. The pLPV plasmids can easily be introduced by electroporation into MDR A. baumannii belonging to the major international lineages as well as into species of the Acinetobacter calcoaceticus-A. baumannii complex. The pLPV vectors have successfully been employed to study the regulation of stress-responsive A. baumannii promoters, including the DNA damage-inducible uvrABC promoter, the ethanol-inducible adhP and yahK promoters, and the iron-repressible promoter of the acinetobactin siderophore biosynthesis gene basA. A lux-tagged A. baumannii ATCC 19606T strain, carrying the iron-responsive pLPV1Z::PbasA promoter fusion, allowed in vivo and ex vivo monitoring of the bacterial burden in the Galleria mellonella infection model. IMPORTANCE The short-term adaptive response to environmental cues greatly contributes to the ecological success of bacteria, and profound alterations in bacterial gene expression occur in response to physical, chemical, and nutritional stresses. Bacteria belonging to the Acinetobacter genus are ubiquitous inhabitants of soil and water though some species, such as Acinetobacter baumannii, are pathogenic and cause serious concern due to antibiotic resistance. Understanding A. baumannii pathobiology requires adequate genetic tools for gene expression analysis, and to this end we developed user-friendly shuttle vectors to probe the transcriptional responses to different environmental stresses. Vectors were constructed to overcome the problem of antibiotic selection in multidrug-resistant strains and were equipped with suitable reporter systems to facilitate signal detection. By means of these vectors, the transcriptional response of A. baumannii to DNA damage, ethanol exposure, and iron starvation was investigated both in vitro and in vivo, providing insights into A. baumannii adaptation during stress and infection.

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129 POST-HATCHING DEVELOPMENT AND GENE EXPRESSION OF IN VITRO-PRODUCED BOVINE EMBRYOS IN RECIPIENT UTERUS AFTER MULTIPLE TRANSFER

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab129 ◽

2013 ◽

Vol 25 (1) ◽

pp. 212

Author(s):

G. Machado ◽

A. Ferreira ◽

I. Pivato ◽

A. Fidelis ◽

J. F. Srpicigo ◽

...

Keyword(s):

Gene Expression ◽

Expression Analysis ◽

Gene Expression Analysis ◽

T Test ◽

Molecular Profile ◽

Control Group ◽

Fluid Medium ◽

Uterine Flushing

This study aimed to compare post-hatching development of Day 7 in vitro and in vivo embryos cultured in recipient uterus until Day 14. For producing in vitro embryos (IVP), oocytes were matured, fertilized (Day 0) and cultured in vitro for 6 days (Day 7) in synthetic oviduct fluid medium supplemented with 5% of fetal bovine serum and incubated at 39°C in 5% CO2 in air. At Day 7, part of IVP blastocysts was transferred to recipient uterus and part was stored for gene expression analysis. As a control group, in vivo embryos were produced after ovarian stimulation, insemination and uterine flushing on Day 7 post insemination. Similarly to the IVP embryos, part of embryos was transferred to recipient uterus and part was stored for gene expression analysis. Day 7 in vivo (n = 53) and IVP (n = 64) expanded blastocysts were transferred to synchronized recipients (10/horn) and were collected by uterine flushing 7 days after transfer (Day 14). Recovered embryos were measured using Motic Image Plus software and evaluated for presence and size of embryonic disc (ED). A trophoblast biopsy was removed and stored for gene expression analysis. For the molecular profile evaluation of Day 7 and Day 14 in vivo and in vitro embryos, 8 genes related with placentation, implantation, oxidative stress, and glucose metabolism (PLAC8, CD9, GLUT-1, GLUT-3, KRT8, MnSOD, HSP70, and INFT, respectively) were quantified by RT-qPCR using ΔΔCT method and CYC-A gene as endogenous control. The recovery rate of Day 14 embryos, analyzed by chi-square test, was higher (P < 0.05) for in vitro than for in vivo embryos, being 50.0% (64/128) and 38.6% (53/137), respectively. No differences (P > 0.05; t-test) were observed in embryo length when comparing Day 14 in vitro (19.1 ± 2.4 mm) and in vivo embryos (24.2 ± 3.7 mm). ED was detected in 25% (16/64) of in vitro and in 26% (14/53) of in vivo embryos. No differences were found (P > 0.05; t-test) in diameter between the two types of embryos (0.3 ± 0.0 mm/in vitro and 0.3 ± 0.0 mm/in vivo). Regarding gene expression, Day 7 IVP embryos showed higher (P < 0.05, Mann–Whitney test) expression of HSP70 and SCL2A1 than in vivo embryos. However, at Day 14 no differences between embryos were observed in transcript levels for any of the studied genes. Therefore, the present study showed that although differences in Day 7 in vitro embryos were observed at the molecular level compared to in vivo counterpart, after transfer to the uterine environment, they showed similar morphology and gene expression profile. These results highlight the importance of evaluating embryos produced by assisted reproductive techniques in later stages of development to have a more precise evaluation of their quality. Financial support: Embrapa, CNPq, CAPES.

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