An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis

Bunnag, Nawat; Tan, Qian; Kaur, Prameet; Ramamoorthy, Anupriya; Sung, Isabelle Chiao Han; Lusk, Jay B; Tolwinski, Nicholas S.

doi:10.1016/j.jmb.2020.03.019

Cited by 16 publications

(15 citation statements)

References 79 publications

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“…As these are adult tissues, genetic approaches must circumvent developmental stages using temperature-sensitive or tissue-specific overexpression, knockdown, or CRISPR/Cas9 [ 87 – 89 ]. One approach used signalling pathway components fused to CRY2 to regulate signalling using light in the adult fly midgut stem cell model [ 16 ]. A limitation of traditional overexpression or knockout techniques is that they affect stem cell regulation across the entire lifespan of the fly.…”

Section: Optogenetic Approachesmentioning

confidence: 99%

“…It will begin transcription of target genes that contain the antioxidant response element (ARE) motif. a Potential optogenetic perturbation of Keap1 (with a fused optogenetic construct such as CRY2/CIB) could induce clustering [ 16 , 137 ] and prevent Keap1 from forming a homodimer and functioning normally. This perturbation could act as a potential gain-of-function of Nrf2 target gene transcription.…”

Section: Optogenetic Approachesmentioning

confidence: 99%

“…As summarized in Table 1 , stem cell signalling pathways that have already been studied using optogenetics include Wnt [ 137 ], RTK (receptor tyrosine kinase) [ 138 ], Ras/ERK (extracellular signal regulated kinase) [ 22 ], Toll, EGFR (epidermal growth factor receptor) [ 16 , 139 ], BMP (bone morphogenetic protein) [ 140 ], and FGF (fibroblast growth factor) [ 141 ]. Conversely, a non-exhaustive list of other potential signalling pathways which have not yet been studied with optogenetics (at the time of this review), but have significant potential include Notch, Hedgehog, Hippo/YAP (Yes-associated protein), and JAK/STAT (Janus kinase/signal transducers and activators of transcription) (Table 2 ).…”

Section: Optogenetic Approachesmentioning

confidence: 99%

“…To manipulate genes directly, without overexpression, many genes in the Drosophila genome have been modified with a landing site for recombination-mediated cassette exchange (RMCE) [ 14 , 15 ]. This landing site can be used to introduce any tag as an additional exon allowing expression of a tagged version of the protein at endogenous levels, and has been used to introduce an optogenetic cassette directly into the genome [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

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Optogenetic approaches for understanding homeostatic and degenerative processes in Drosophila

Lim

Kaur

Huang

et al. 2021

Cell. Mol. Life Sci.

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Many organs and tissues have an intrinsic ability to regenerate from a dedicated, tissue-specific stem cell pool. As organisms age, the process of self-regulation or homeostasis begins to slow down with fewer stem cells available for tissue repair. Tissues become more fragile and organs less efficient. This slowdown of homeostatic processes leads to the development of cellular and neurodegenerative diseases. In this review, we highlight the recent use and future potential of optogenetic approaches to study homeostasis. Optogenetics uses photosensitive molecules and genetic engineering to modulate cellular activity in vivo , allowing precise experiments with spatiotemporal control. We look at applications of this technology for understanding the mechanisms governing homeostasis and degeneration as applied to widely used model organisms, such as Drosophila melanogaster , where other common tools are less effective or unavailable.

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Section: Optogenetic Approachesmentioning

confidence: 99%

Section: Optogenetic Approachesmentioning

confidence: 99%

Section: Optogenetic Approachesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Optogenetic approaches for understanding homeostatic and degenerative processes in Drosophila

Lim

Kaur

Huang

et al. 2021

Cell. Mol. Life Sci.

Self Cite

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“…We and others have recently engineered light-activated variants of key signaling proteins that now provide a basis for the optogenetic delivery of trophic signals. Particular success was reported for receptor tyrosine kinases (RTKs) (14)(15)(16)(17)(18). RTKs are expressed in virtually all human cell types and respond to growth factors (GFs) with conformational changes and/or oligomerization state changes that result in receptor trans-phosphorylation.…”

Section: Introductionmentioning

confidence: 99%

Optogenetic delivery of trophic signals in a genetic model of Parkinson’s disease

Inglés-Prieto

Furthmann

Crossman

et al. 2020

Preprint

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Optogenetics has been harnessed to shed new mechanistic light on current therapies and to develop future treatment strategies. This has been to date achieved by the correction of electrical signals in neuronal cells and neural circuits that are affected by disease. In contrast, the optogenetic delivery of trophic biochemical signals, which support cell survival and thereby may modify progression of degenerative disorders, has never been demonstrated in an animal disease model. Here, we reengineered the human and Drosophila melanogaster REarranged during Transfection (hRET and dRET) receptors to be activated by light, creating one-component optogenetic tools termed Opto-hRET and Opto-dRET. Upon blue light stimulation, these receptors robustly induced the MAPK/ERK proliferative signaling pathway in cultured cells. In PINK1B9 flies that exhibit loss of PTEN-induced putative kinase 1 (PINK1), a kinase associated with familial Parkinson’s disease (PD), light activation of Opto-dRET suppressed mitochondrial defects, tissue degeneration and behavioral deficits. In human cells with PINK1 loss-of-function, mitochondrial fragmentation was rescued using Opto-dRET via the PI3K/NF-кB pathway. Our results demonstrate that a light-activated receptor can ameliorate disease hallmarks in a genetic model of PD. The optogenetic delivery of trophic signals is cell type-specific and reversible and thus has the potential to overcome limitations of current strategies towards a spatio-temporal regulation of tissue repair.Significance StatementThe death of physiologically important cell populations underlies of a wide range of degenerative disorders, including Parkinson’s disease (PD). Two major strategies to counter cell degeneration, soluble growth factor injection and growth factor gene therapy, can lead to the undesired activation of bystander cells and non-natural permanent signaling responses. Here, we employed optogenetics to deliver cell type-specific pro-survival signals in a genetic model of PD. In Drosophila and human cells exhibiting loss of the PINK1 kinase, akin to autosomal recessive PD, we efficiently suppressed disease phenotypes using a light-activated tyrosine kinase receptor. This work demonstrates a spatio-temporally precise strategy to interfere with degeneration and may open new avenues towards tissue repair in disease models.

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The early Drosophila embryo as a model system for quantitative biology

Saunders

2021

Cells & Development

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An Optogenetic Method to Study Signal Transduction in Intestinal Stem Cell Homeostasis

Cited by 16 publications

References 79 publications

Optogenetic approaches for understanding homeostatic and degenerative processes in Drosophila

Optogenetic approaches for understanding homeostatic and degenerative processes in Drosophila

Optogenetic delivery of trophic signals in a genetic model of Parkinson’s disease

The early Drosophila embryo as a model system for quantitative biology

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