Life Sciences Research for Lifelong Health

Oliver Florey

Research Summary

Research in our lab is focused on the related topics of autophagy (self eating), macroendocytosis (digestion of extracellular material) and entosis (a recently discovered form of cell cannibalism). These are 3 distinct but inter-related forms of cellular ‘eating’, which play an important role in normal biology and become deregulated during ageing or disease (eg cancer).

Our work exploits a combination of molecular and cellular biology, state-of-the-art microscopy (long-term timelapse imaging, spinning disk confocal and electron microscopy) and proteomics (mass spectrometry).

Existing projects aim to define the molecular mechanisms which underlie cellular eating, with a particular focus on the emerging pathway of non-canonical autophagy. We are also investigating the intriguing relationship between entosis and cancer.

Latest Publications

Imaging Noncanonical Autophagy and LC3-Associated Phagocytosis in Cultured Cells.
Jacquin E, Fletcher K, Florey O

Monitoring of ATG8 proteins by western blotting and immunofluorescence microscopy are the most common methods to monitor the autophagy pathway. However, it has recently been shown that ATG8 proteins can be lipidated to non-autophagosome, single-membrane compartments through a noncanonical autophagy pathway. This is commonly found to occur during macro-endocytic processes such as phagocytosis, where it has been termed LC3-associated phagocytosis, and upon lysosomotropic drug treatment. Therefore, care is required when interpreting data based on ATG8 in order to conclude whether a signal relates to the canonical or noncanonical pathway. Here we provide methods to monitor noncanonical autophagy through fluorescence microscopy.

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Methods in molecular biology (Clifton, N.J.), 1880, 1940-6029, 295-303, 2019

PMID: 30610705

The double life of autophagy proteins.
Florey O

Nature microbiology, 3, 2058-5276, 1334-1335, 2018

PMID: 30478385

Alpha-synuclein fibrils recruit TBK1 and OPTN to lysosomal damage sites and induce autophagy in microglial cells.
Bussi C, Peralta Ramos JM, Arroyo DS, Gallea JI, Ronchi P, Kolovou A, Wang JM, Florey O, Celej MS, Schwab Y, Ktistakis NT, Iribarren P

Autophagic dysfunction and protein aggregation have been linked to several neurodegenerative disorders, but the exact mechanisms and causal connections are not clear and most work was done in neurons and not in microglial cells. Here we report that exogenous fibrillar but not monomeric alpha-synuclein (AS) induces autophagy in microglial cells. We extensively studied the dynamics of this response by both live-cell imaging and correlative light-electron microscopy (CLEM) and found that it correlates with lysosomal damage and is characterised by the recruitment of the selective autophagy-associated proteins TANK-binding kinase 1 (TBK1) and Optineurin (OPTN) to ubiquitinated lysosomes. In addition, we observed that LC3 recruitment to damaged lysosomes was dependent on TBK1 activity. In these fibrillar AS-treated cells, autophagy inhibition impairs mitochondrial function and leads to microglial cell death. Our results suggest that microglial autophagy is induced in response to lysosomal damage caused by persistent accumulation of AS fibrils. Importantly, triggering of the autophagic response appears to be an attempt at lysosomal quality control and not for engulfment of fibrillar AS.

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Journal of cell science, , 1477-9137, , 2018

PMID: 30404831

entosis and the formation of a cell-in-cell structure by MCF10A cells
A) Sequence of images showing entosis and the formation of a cell-in-cell structure by MCF10A cells in suspension. Cell 1 is engulfed by Cell 2.

B) H&E staining from a human breast carcinoma, arrows point to cell-in-cell structures (taken from Biomax.us).

C) Immunofluorescent staining of b-catenin in a cell-in-cell structure from a human breast tumor.

D) Immunofluorescent staining of E-cadherin in a cell-in-cell structure from MCF10A cells.

Group Members

Latest Publications

Imaging Noncanonical Autophagy and LC3-Associated Phagocytosis in Cultured Cells.

Jacquin E, Fletcher K, Florey O

Methods in molecular biology (Clifton, N.J.)
1880 1940-6029:295-303 (2019)

PMID: 30610705

The double life of autophagy proteins.

Florey O

Nature microbiology
3 2058-5276:1334-1335 (2018)

PMID: 30478385

Alpha-synuclein fibrils recruit TBK1 and OPTN to lysosomal damage sites and induce autophagy in microglial cells.

Bussi C, Peralta Ramos JM, Arroyo DS

Journal of cell science
1477-9137: (2018)

PMID: 30404831

Mitosis can drive cell cannibalism through entosis.

Durgan J, Tseng YY, Hamann JC

eLife
6 2050-084X: (2017)

PMID: 28693721

PIKfyve Regulates Vacuole Maturation and Nutrient Recovery following Engulfment.

Krishna S, Palm W, Lee Y

Developmental cell
38 1878-1551:536-47 (2016)

PMID: 27623384

3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy.

Russell MR, Lerner TR, Burden JJ

Journal of cell science
1477-9137: (2016)

PMID: 27445312

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).

Klionsky DJ, Abdelmohsen K, Abe A

Autophagy
12 1554-8635:1-222 (2016)

PMID: 26799652

V-ATPase and osmotic imbalances activate endolysosomal LC3 lipidation.

Florey O, Gammoh N, Kim SE

Autophagy
1554-8635:0 (2014)

PMID: 25484071

SOS1 and Ras regulate epithelial tight junction formation in the human airway through EMP1.

Durgan J, Tao G, Walters MS

EMBO reports
16 1469-3178:87-96 (2015)

PMID: 25394671