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Article ; Online: Tightrope act: autophagy in stem cell renewal, differentiation, proliferation, and aging.

Phadwal, Kanchan / Watson, Alexander Scarth / Simon, Anna Katharina

Cellular and molecular life sciences : CMLS

2012 Volume 70, Issue 1, Page(s) 89–103

Abstract: Autophagy is a constitutive lysosomal catabolic pathway that degrades damaged organelles and protein aggregates. Stem cells are characterized by self-renewal, pluripotency, and quiescence; their long life span, limited capacity to dilute cellular waste ... ...

Abstract	Autophagy is a constitutive lysosomal catabolic pathway that degrades damaged organelles and protein aggregates. Stem cells are characterized by self-renewal, pluripotency, and quiescence; their long life span, limited capacity to dilute cellular waste and spent organelles due to quiescence, along with their requirement for remodeling in order to differentiate, all suggest that they require autophagy more than other cell types. Here, we review the current literature on the role of autophagy in embryonic and adult stem cells, including hematopoietic, mesenchymal, and neuronal stem cells, highlighting the diverse and contrasting roles autophagy plays in their biology. Furthermore, we review the few studies on stem cells, lysosomal activity, and autophagy. Novel techniques to detect autophagy in primary cells are required to study autophagy in different stem cell types. These will help to elucidate the importance of autophagy in stem cells during transplantation, a promising therapeutic approach for many diseases.
MeSH term(s)	Animals ; Autophagy/physiology ; Cell Differentiation ; Cell Proliferation ; Cellular Senescence ; Humans ; Lysosomes/physiology ; Mice ; Models, Biological ; Stem Cell Transplantation ; Stem Cells/cytology ; Stem Cells/physiology
Language	English
Publishing date	2012-06-05
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-012-1032-3
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Article ; Online: Autophagy in the pathogenesis of myelodysplastic syndrome and acute myeloid leukemia.

Watson, Alexander Scarth / Mortensen, Monika / Simon, Anna Katharina

Cell cycle (Georgetown, Tex.)

2011 Volume 10, Issue 11, Page(s) 1719–1725

Abstract: Autophagy is a conserved cellular pathway responsible for the sequestration of spent organelles and protein aggregates from the cytoplasm and their delivery into lysosomes for degradation. Autophagy plays an important role in adaptation to starvation, in ...

Abstract	Autophagy is a conserved cellular pathway responsible for the sequestration of spent organelles and protein aggregates from the cytoplasm and their delivery into lysosomes for degradation. Autophagy plays an important role in adaptation to starvation, in cell survival, immunity, development and cancer. Recent evidence in mice suggests that autophagic defects in hematopoietic stem cells (HSCs) may be implicated in leukemia. Indeed, mice lacking Atg7 in HSCs develop an atypical myeloproliferation resembling human myelodysplastic syndrome (MDS) progressing to acute myeloid leukemia (AML). Studies suggest that accumulation of damaged mitochondria and reactive oxygen species result in cell death of the majority of progenitor cells and, possibly, concomitant transformation of some surviving ones. Interestingly, bone marrow cells from MDS patients are characterized by mitochondrial abnormalities and increased cell death. A role for autophagy in the transformation to cancer has been proposed in other cancer types. This review focuses on autophagy in human MDS development and progression to AML within the context of the role of mitochondria, apoptosis and reactive oxygen species (ROS) in its pathogenesis.
MeSH term(s)	Autophagy/physiology ; Bone Marrow Cells/pathology ; Bone Marrow Cells/ultrastructure ; Humans ; Leukemia, Myeloid, Acute/etiology ; Leukemia, Myeloid, Acute/pathology ; Mitochondria ; Myelodysplastic Syndromes/etiology ; Myelodysplastic Syndromes/pathology ; Reactive Oxygen Species
Chemical Substances	Reactive Oxygen Species
Language	English
Publishing date	2011-06-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2146183-1
ISSN	1551-4005 ; 1538-4101 ; 1554-8627
ISSN (online)	1551-4005
ISSN	1538-4101 ; 1554-8627
DOI	10.4161/cc.10.11.15673
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Article ; Online: Lack of autophagy in the hematopoietic system leads to loss of hematopoietic stem cell function and dysregulated myeloid proliferation.

Mortensen, Monika / Watson, Alexander Scarth / Simon, Anna Katharina

Autophagy

2011 Volume 7, Issue 9, Page(s) 1069–1070

Abstract: The regulated lysosomal degradation pathway of autophagy prevents cellular damage and thus protects from malignant transformation. Autophagy is also required for the maturation of various hematopoietic lineages, namely the erythroid and lymphoid ones, ... ...

Abstract	The regulated lysosomal degradation pathway of autophagy prevents cellular damage and thus protects from malignant transformation. Autophagy is also required for the maturation of various hematopoietic lineages, namely the erythroid and lymphoid ones, yet its role in adult hematopoietic stem cells (HSCs) remained unexplored. While normal HSCs sustain life-long hematopoiesis, malignant transformation of HSCs or early progenitors leads to leukemia. Mechanisms protecting HSCs from cellular damage are therefore essential to prevent hematopoietic malignancies. By conditionally deleting the essential autophagy gene Atg7 in the hematopoietic system, we found that autophagy is required for the maintenance of true HSCs and therefore also of downstream hematopoietic progenitors. Loss of autophagy in HSCs leads to the expansion of a progenitor cell population in the bone marrow, giving rise to a severe, invasive myeloproliferation, which strongly resembles human acute myeloid leukemia (AML).
MeSH term(s)	Animals ; Autophagy ; Autophagy-Related Protein 7 ; Cell Proliferation ; Hematopoietic Stem Cells/metabolism ; Hematopoietic Stem Cells/pathology ; Hematopoietic System/metabolism ; Hematopoietic System/pathology ; Mice ; Microtubule-Associated Proteins/deficiency ; Microtubule-Associated Proteins/metabolism ; Myeloid Cells/metabolism ; Myeloid Cells/pathology
Chemical Substances	Atg7 protein, mouse ; Microtubule-Associated Proteins ; Autophagy-Related Protein 7 (EC 6.2.1.45)
Language	English
Publishing date	2011-09-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	2454135-7
ISSN	1554-8635 ; 1554-8627
ISSN (online)	1554-8635
ISSN	1554-8627
DOI	10.4161/auto.7.9.15886
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Article ; Online: Techniques for the Detection of Autophagy in Primary Mammalian Cells.

Puleston, Daniel / Phadwal, Kanchan / Watson, Alexander Scarth / Soilleux, Elizabeth J / Chittaranjan, Suganthi / Bortnik, Svetlana / Gorski, Sharon M / Ktistakis, Nicholas / Simon, Anna Katharina

Cold Spring Harbor protocols

2015 Volume 2015, Issue 9, Page(s) pdb.top070391

Abstract: Autophagy is a lysosomal catabolic pathway responsible for the degradation of cytoplasmic constituents. Autophagy is primarily a survival pathway for recycling cellular material in times of nutrient starvation, and in response to hypoxia, endoplasmic ... ...

Abstract	Autophagy is a lysosomal catabolic pathway responsible for the degradation of cytoplasmic constituents. Autophagy is primarily a survival pathway for recycling cellular material in times of nutrient starvation, and in response to hypoxia, endoplasmic reticulum stress, and other stresses, regulated through the mammalian target of rapamycin pathway. The proteasomal pathway is responsible for degradation of proteins, whereas autophagy can degrade cytoplasmic material in bulk, including whole organelles such as mitochondria (mitophagy), bacteria (xenophagy), or lipids (lipophagy). Although signs of autophagy can be present during cell death, it remains controversial whether autophagy can execute cell death in vivo. Here, we will introduce protocols for detecting autophagy in mammalian primary cells by using western blots, immunofluorescence, immunohistochemistry, flow cytometry, and imaging flow cytometry.
MeSH term(s)	Animals ; Autophagy ; Cell Physiological Phenomena ; Cells, Cultured ; Cytological Techniques/methods ; Humans ; Mammals
Language	English
Publishing date	2015-09-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1559-6095
ISSN (online)	1559-6095
DOI	10.1101/pdb.top070391
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Article ; Online: Autophagy Controls Acquisition of Aging Features in Macrophages.

Stranks, Amanda J / Hansen, Anne Louise / Panse, Isabel / Mortensen, Monika / Ferguson, David J P / Puleston, Daniel J / Shenderov, Kevin / Watson, Alexander Scarth / Veldhoen, Marc / Phadwal, Kanchan / Cerundolo, Vincenzo / Simon, Anna Katharina

Journal of innate immunity

2015 Volume 7, Issue 4, Page(s) 375–391

Abstract: Macrophages provide a bridge linking innate and adaptive immunity. An increased frequency of macrophages and other myeloid cells paired with excessive cytokine production is commonly seen in the aging immune system, known as 'inflamm-aging'. It is ... ...

Abstract	Macrophages provide a bridge linking innate and adaptive immunity. An increased frequency of macrophages and other myeloid cells paired with excessive cytokine production is commonly seen in the aging immune system, known as 'inflamm-aging'. It is presently unclear how healthy macrophages are maintained throughout life and what connects inflammation with myeloid dysfunction during aging. Autophagy, an intracellular degradation mechanism, has known links with aging and lifespan extension. Here, we show for the first time that autophagy regulates the acquisition of major aging features in macrophages. In the absence of the essential autophagy gene Atg7, macrophage populations are increased and key functions such as phagocytosis and nitrite burst are reduced, while the inflammatory cytokine response is significantly increased - a phenotype also observed in aged macrophages. Furthermore, reduced autophagy decreases surface antigen expression and skews macrophage metabolism toward glycolysis. We show that macrophages from aged mice exhibit significantly reduced autophagic flux compared to young mice. These data demonstrate that autophagy plays a critical role in the maintenance of macrophage homeostasis and function, regulating inflammation and metabolism and thereby preventing immunosenescence. Thus, autophagy modulation may prevent excess inflammation and preserve macrophage function during aging, improving immune responses and reducing the morbidity and mortality associated with inflamm-aging.
MeSH term(s)	Aging/genetics ; Aging/immunology ; Aging/pathology ; Animals ; Autophagy/genetics ; Autophagy/immunology ; Autophagy-Related Protein 7 ; Glycolysis/genetics ; Glycolysis/immunology ; Macrophages/immunology ; Macrophages/pathology ; Mice ; Mice, Knockout ; Microtubule-Associated Proteins/genetics ; Microtubule-Associated Proteins/immunology
Chemical Substances	Atg7 protein, mouse ; Microtubule-Associated Proteins ; Autophagy-Related Protein 7 (EC 6.2.1.45)
Language	English
Publishing date	2015-03-10
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2454158-8
ISSN	1662-8128 ; 1662-811X
ISSN (online)	1662-8128
ISSN	1662-811X
DOI	10.1159/000370112
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Article ; Online: p38 signaling inhibits mTORC1-independent autophagy in senescent human CD8⁺ T cells.

Henson, Sian M / Lanna, Alessio / Riddell, Natalie E / Franzese, Ornella / Macaulay, Richard / Griffiths, Stephen J / Puleston, Daniel J / Watson, Alexander Scarth / Simon, Anna Katharina / Tooze, Sharon A / Akbar, Arne N

The Journal of clinical investigation

2014 Volume 124, Issue 9, Page(s) 4004–4016

Abstract: T cell senescence is thought to contribute to immune function decline, but the pathways that mediate senescence in these cells are not clear. Here, we evaluated T cell populations from healthy volunteers and determined that human CD8+ effector memory T ... ...

Abstract	T cell senescence is thought to contribute to immune function decline, but the pathways that mediate senescence in these cells are not clear. Here, we evaluated T cell populations from healthy volunteers and determined that human CD8+ effector memory T cells that reexpress the naive T cell marker CD45RA have many characteristics of cellular senescence, including decreased proliferation, defective mitochondrial function, and elevated levels of both ROS and p38 MAPK. Despite their apparent senescent state, we determined that these cells secreted high levels of both TNF-α and IFN-γ and showed potent cytotoxic activity. We found that the senescent CD45RA-expressing population engaged anaerobic glycolysis to generate energy for effector functions. Furthermore, inhibition of p38 MAPK signaling in senescent CD8+ T cells increased their proliferation, telomerase activity, mitochondrial biogenesis, and fitness; however, the extra energy required for these processes did not arise from increased glucose uptake or oxidative phosphorylation. Instead, p38 MAPK blockade in these senescent cells induced an increase in autophagy through enhanced interactions between p38 interacting protein (p38IP) and autophagy protein 9 (ATG9) in an mTOR-independent manner. Together, our findings describe fundamental metabolic requirements of senescent primary human CD8+ T cells and demonstrate that p38 MAPK blockade reverses senescence via an mTOR-independent pathway.
MeSH term(s)	Adult ; Autophagy/physiology ; Autophagy-Related Proteins ; CD8-Positive T-Lymphocytes/physiology ; Cellular Senescence ; DNA Damage ; Glycolysis ; Humans ; MAP Kinase Signaling System/physiology ; Mechanistic Target of Rapamycin Complex 1 ; Membrane Potential, Mitochondrial ; Membrane Proteins/physiology ; Mitochondria/physiology ; Multiprotein Complexes/physiology ; Oxidative Phosphorylation ; TOR Serine-Threonine Kinases/physiology ; Vesicular Transport Proteins/physiology ; p38 Mitogen-Activated Protein Kinases/physiology
Chemical Substances	Atg9A protein, mouse ; Autophagy-Related Proteins ; Membrane Proteins ; Multiprotein Complexes ; Vesicular Transport Proteins ; Mechanistic Target of Rapamycin Complex 1 (EC 2.7.11.1) ; TOR Serine-Threonine Kinases (EC 2.7.11.1) ; p38 Mitogen-Activated Protein Kinases (EC 2.7.11.24)
Language	English
Publishing date	2014-08-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI75051
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Article ; Online: A novel method for autophagy detection in primary cells: impaired levels of macroautophagy in immunosenescent T cells.

Phadwal, Kanchan / Alegre-Abarrategui, Javier / Watson, Alexander Scarth / Pike, Luke / Anbalagan, Selvakumar / Hammond, Ester M / Wade-Martins, Richard / McMichael, Andrew / Klenerman, Paul / Simon, Anna Katharina

Autophagy

2012 Volume 8, Issue 4, Page(s) 677–689

Abstract: Autophagy is a conserved constitutive cellular process, responsible for the degradation of dysfunctional proteins and organelles. Autophagy plays a role in many diseases such as neurodegeneration and cancer; however, to date, conventional autophagy ... ...

Abstract	Autophagy is a conserved constitutive cellular process, responsible for the degradation of dysfunctional proteins and organelles. Autophagy plays a role in many diseases such as neurodegeneration and cancer; however, to date, conventional autophagy detection techniques are not suitable for clinical samples. We have developed a high throughput, statistically robust technique that quantitates autophagy in primary human leukocytes using the Image stream, an imaging flow cytometer. We validate this method on cell lines and primary cells knocked down for essential autophagy genes. Also, using this method we show that T cells have higher autophagic activity than B cells. Furthermore our results indicate that healthy primary senescent CD8(+) T cells have decreased autophagic levels correlating with increased DNA damage, which may explain features of the senescent immune system and its declining function with age. This technique will allow us, for the first time, to measure autophagy levels in diseases with a known link to autophagy, while also determining the contribution of autophagy to the efficacy of drugs.
MeSH term(s)	Adult ; Animals ; Autophagy/immunology ; B-Lymphocytes/cytology ; B-Lymphocytes/immunology ; Biological Assay/methods ; Cells, Cultured ; Cellular Senescence/immunology ; HEK293 Cells ; Histones/metabolism ; Humans ; Mice ; Middle Aged ; Reproducibility of Results ; T-Lymphocytes/cytology ; T-Lymphocytes/immunology
Chemical Substances	H2AX protein, human ; Histones
Language	English
Publishing date	2012-04-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2454135-7
ISSN	1554-8635 ; 1554-8627
ISSN (online)	1554-8635
ISSN	1554-8627
DOI	10.4161/auto.18935
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Article: Autophagy Controls Acquisition of Aging Features in Macrophages

Stranks, Amanda J. / Hansen, Anne Louise / Panse, Isabel / Mortensen, Monika / Ferguson, David J.P. / Puleston, Daniel J. / Shenderov, Kevin / Watson, Alexander Scarth / Veldhoen, Marc / Phadwal, Kanchan / Cerundolo, Vincenzo / Simon, Anna Katharina

Journal of Innate Immunity

2015 Volume 7, Issue 4, Page(s) 375–391

Institution	MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford BRC Translational Immunology Lab, Experimental Medicine, Nuffield Department of Medicine, and Nuffield Department of Clinical and Laboratory Sciences, John Radcliffe Hospital, Oxford Babraham Institute, Cambridge, UK Cell Death and Metabolism, Danish Cancer Society Research Center, Danish Cancer Society, Copenhagen, Denmark Johns Hopkins University School of Medicine, Baltimore, Md., USA
Abstract	Macrophages provide a bridge linking innate and adaptive immunity. An increased frequency of macrophages and other myeloid cells paired with excessive cytokine production is commonly seen in the aging immune system, known as ‘inflamm-aging'. It is presently unclear how healthy macrophages are maintained throughout life and what connects inflammation with myeloid dysfunction during aging. Autophagy, an intracellular degradation mechanism, has known links with aging and lifespan extension. Here, we show for the first time that autophagy regulates the acquisition of major aging features in macrophages. In the absence of the essential autophagy gene Atg7, macrophage populations are increased and key functions such as phagocytosis and nitrite burst are reduced, while the inflammatory cytokine response is significantly increased - a phenotype also observed in aged macrophages. Furthermore, reduced autophagy decreases surface antigen expression and skews macrophage metabolism toward glycolysis. We show that macrophages from aged mice exhibit significantly reduced autophagic flux compared to young mice. These data demonstrate that autophagy plays a critical role in the maintenance of macrophage homeostasis and function, regulating inflammation and metabolism and thereby preventing immunosenescence. Thus, autophagy modulation may prevent excess inflammation and preserve macrophage function during aging, improving immune responses and reducing the morbidity and mortality associated with inflamm-aging.
Keywords	Autophagy ; Macrophages ; Aging ; Inflamm-aging ; Inflammation ; Metabolism
Language	English
Publishing date	2015-03-10
Publisher	S. Karger AG
Publishing place	Basel, Switzerland
Document type	Article
ZDB-ID	2454158-8
ISSN	1662-8128 ; 1662-811X
ISSN (online)	1662-8128
ISSN	1662-811X
DOI	10.1159/000370112
Database	Karger publisher's database

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Article: Cross‐cultural collaboration leads to greater understanding of the rare Spectacled Hare‐wallaby in the west Kimberley, Western Australia

Wysong, Michael L. / Gregory, Pius / Watson, Alexander W. T. / Woolley, Leigh‐Ann / Parker, Christopher W. / Country Managers, Yawuru / Rangers, Karajarri / Mangala Rangers, Nyikina

Ecological management & restoration. 2022 Jan., v. 23 Suppl S1

2022

Abstract: ... a negative relationship between SHW occurrence and distance to fire scar edge burnt in current or previous ...

Abstract	Cross‐cultural collaboration between Yawuru Country Managers (Rangers) and WWF‐Australia ecologists led to new detections of the Spectacled Hare‐wallaby (SHW), (Lagorchestes conspicillatus) in the west Kimberley region of Western Australia where it was presumed to be locally extirpated. This collaboration relied on the expertise of the Yawuru Country Managers to select specific locations for targeted field surveys and resulted in the confirmation of SHW on the Yawuru IPA for the first time in a decade. Subsequent remote camera trap surveys over a larger area included collaboration with two additional neighbouring Indigenous ranger groups, Karrajarri and Nyikina Mangala. These surveys investigated the spatial and temporal relationship between SHW and other mammals which may threaten (e.g., feral Cat [Felis catus], Dingo [Canis familiaris dingo]) or compete (e.g., Agile Wallaby [Macropus agilis]; Cattle [Bos taurus]) with them. We found a negative relationship between SHW and cat activity, suggesting that cats may limit the activity or abundance of SHW. Temporal portioning was evident between SHW and both Cattle and Agile Wallaby suggesting that SHW may avoid times when these species are most active. Further, we found a negative relationship between SHW occurrence and distance to fire scar edge burnt in current or previous fire season. This edge habitat is likely important to SHW because they may require recently burnt areas to forage and dense unburnt areas to shelter. This project highlights the benefits of cross‐cultural research and monitoring partnerships with Indigenous rangers as active observers and managers of their traditional lands.
Keywords	Lagorchestes ; Macropus agilis ; administrative management ; cameras ; cats ; cattle ; dingoes ; edge effects ; fire season ; Western Australia
Language	English
Dates of publication	2022-01
Size	p. 139-149.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	2020194-1
ISSN	1442-8903 ; 1442-7001
ISSN (online)	1442-8903
ISSN	1442-7001
DOI	10.1111/emr.12524
Database	NAL-Catalogue (AGRICOLA)

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Article: Tightrope act: autophagy in stem cell renewal, differentiation, proliferation, and aging

Phadwal, Kanchan / Watson, Alexander Scarth / Simon, Anna Katharina

Cellular and molecular life sciences

Volume v. 70,, Issue no. 1

Abstract	Autophagy is a constitutive lysosomal catabolic pathway that degrades damaged organelles and protein aggregates. Stem cells are characterized by self-renewal, pluripotency, and quiescence; their long life span, limited capacity to dilute cellular waste and spent organelles due to quiescence, along with their requirement for remodeling in order to differentiate, all suggest that they require autophagy more than other cell types. Here, we review the current literature on the role of autophagy in embryonic and adult stem cells, including hematopoietic, mesenchymal, and neuronal stem cells, highlighting the diverse and contrasting roles autophagy plays in their biology. Furthermore, we review the few studies on stem cells, lysosomal activity, and autophagy. Novel techniques to detect autophagy in primary cells are required to study autophagy in different stem cell types. These will help to elucidate the importance of autophagy in stem cells during transplantation, a promising therapeutic approach for many diseases.
Keywords	protein aggregates ; organelles ; autophagy ; longevity ; adult stem cells
Language	English
Document type	Article
ISSN	1420-682X
Database	AGRIS - International Information System for the Agricultural Sciences and Technology

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