Harnessing autophagy for cell fate control gene therapy

Felizardo, Tania C.; Foley, Jason; Steed, Kevin; Dropulić, Boro; Amarnath, Shoba; Medin, Jeffrey A.; Fowler, Daniel H.

doi:10.4161/auto.24639

Cited by 8 publications

(6 citation statements)

References 38 publications

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“…We have previously described our rapamycin conditioning protocol to generate a T h 2‐polarized phenotype (Fowler et al , 2013 ). We then integrated a single transduction step to this protocol, previously optimized using a lentivirus that engineers expression of eGFP to consistently obtain at least 40% eGFP‐expressing cells (Felizardo et al , 2013 ), to generate T‐Rapa micropharmacies (TRaMs; Fig 1A ). As part of our internal quality control, and, in addition to the previously studied benefits of rapamycin conditioning (Mariotti et al , 2008 ; He et al , 2011 ; Fowler et al , 2013 ), TRaMs appeared to be more resilient to cryogenic stress (Fig EV2A ) and may have better potential to expand than control T cells produced without rapamycin conditioning (Fig EV2B ).…”

Section: Resultsmentioning

confidence: 99%

“…T‐Rapa were produced, cultured, and transduced essentially as described previously (Felizardo et al , 2013 ). Briefly, T cells were conditioned with 1 µM rapamycin (sirolimus oral solution, Pfizer) for 3 days with the following culture conditions: X‐VIVO 20 media supplemented with 5% v/v human AB serum (Gemini bio‐products 100‐512, various lots), 20 U/ml of interleukin (IL)‐2 (Roche 11147528001), 1,000 U/ml of IL‐4 (R&D Systems 204‐IL), and CD3/CD28 T‐Activator beads (Gibco 111.32D) at a 3:1 bead‐to‐cell ratio.…”

Section: Methodsmentioning

confidence: 99%

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Autologous, lentivirus‐modified, T‐rapa cell “micropharmacies” for lysosomal storage disorders

Nagree

Felizardo²,

Faber

et al. 2022

EMBO Mol Med

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T cells are the current choice for many cell therapy applications. They are relatively easy to access, expand in culture, and genetically modify. Rapamycin-conditioning ex vivo reprograms T cells, increasing their memory properties and capacity for survival, while reducing inflammatory potential and the amount of preparative conditioning required for engraftment. Rapamycinconditioned T cells have been tested in patients and deemed to be safe to administer in numerous settings, with reduced occurrence of infusion-related adverse events. We demonstrate that ex vivo lentivirus-modified, rapamycin-conditioned CD4 + T cells can also act as next-generation cellular delivery vehicles-that is, "micropharmacies"-to disseminate corrective enzymes for multiple lysosomal storage disorders. We evaluated the therapeutic potential of this treatment platform for Fabry, Gaucher, Farber, and Pompe diseases in vitro and in vivo. For example, such micropharmacies expressing a-galactosidase A for treatment of Fabry disease were transplanted in mice where they provided functional enzyme in key affected tissues such as kidney and heart, facilitating clearance of pathogenic substrate after a single administration.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Autologous, lentivirus‐modified, T‐rapa cell “micropharmacies” for lysosomal storage disorders

Nagree

Felizardo²,

Faber

et al. 2022

EMBO Mol Med

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“…Consistent with this, our results showed that the expression of mTOR was increased in APP/PS1 transgenic mice, which may cause the phosphorylation of IRS-1 Ser636 and lead to insulin resistance. 18 Furthermore, HNG treatment downregulated mTOR expression, decreased the phosphorylation of IRS-1 Ser636, and alleviated insulin resistance.…”

Section: Discussionmentioning

confidence: 98%

S14G‐humanin alleviates insulin resistance and increases autophagy in neurons of APP/PS1 transgenic mouse

Han

Jia

Zhong

et al. 2017

J of Cellular Biochemistry

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Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by Aβ plaque deposition in the brain, which is related to the disorder of autophagosome maturation, transport, and formation of autolysosome. Notably, abnormal insulin signaling is connected with cognitive dysfunction in AD. In this study, using APP/PS1 transgenic mice as AD model, we investigated the mechanism by which S14G-humanin (HNG) improved autophagy and insulin signaling in AD brain. Immunohistochemistry was used to determine the levels of mTOR and Aβ deposition, and Western blot analysis was used to determine IRS-1, IRS-1 pSEr636, ULK1, p62, LC3 I/LC3 II protein levels. Our results demonstrated that HNG could improve the learning ability and memory in APP/PS1 transgenic mice, possibly through decreasing IRS-1 Ser636 phosphorylation and mTOR protein expression in the hippocampus, thus improving insulin resistance in the brain. In addition, HNG increased ULK1 expression, decreased p62 and LC3 I/LC3 II protein levels, thus enhancing autophagy and decreasing Aβ deposition in the brain. Taken together, our results suggest that through the regulation of IRS-1/mTOR insulin signaling in the hippocampus, HNG increases the activity of autophagy and decreases Aβ deposition in the brain, and improves learning ability and memory of AD mice.

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“…As we have recently shown (142), T-Rapa cells are amenable to lentiviral-mediated transfer of a human, modified thymidylate kinase (TMPK) transgene that efficiently activates the pro-drug AZT to execute a new cell fate, suicide gene axis (143). Further studies will be required to determine the merits of this approach relative to previously described suicide gene axes (144, 145).…”

Section: Ongoing and Future Directions In Rapamycin-resistant T-cell mentioning

confidence: 99%

Rapamycin‐resistant effector T‐cell therapy

Fowler

2013

Immunological Reviews

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Pharmacologic inhibition of the mechanistic target of rapamycin (mTOR) represents a stress test for tumor cells and T cells. Mechanisms exist that allow cells to survive this stress, including suboptimal target block, alternative signaling pathways, and autophagy. Rapamycin-resistant effector T (T-Rapa) cells have an altered phenotype that associates with increased function. Ex vivo rapamycin, when used in combination with polarizing cytokines and antigen-presenting-cell free costimulation, is a flexible therapeutic approach as polarization to T-helper 1 (Th1)- or Th2-type effectors is possible. Murine T-Rapa cells skewed toward a Th2-type prevented graft rejection and graft-versus-host disease (GVHD) more potently than control Th2 cells and effectively balanced GVHD and graft-versus-tumor (GVT) effects. A phase II clinical trial using low-intensity allogeneic hematopoietic cell transplantation demonstrated that interleukin-4 polarized human T-Rapa cells had a mixed Th2/Th1 phenotype; T-Rapa cell recipients had a balanced Th2/Th1 cytokine profile, conversion of mixed chimerism toward full donor chimerism, and a potentially favorable balance between GVHD and GVT effects. In addition, a phase I clinical trial evaluating autologous T-Rapa cells skewed toward a Th1- and Tc1-type is underway. Use of ex vivo rapamycin to modulate effector T-cell function represents a promising new approach to transplantation therapy.

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Harnessing autophagy for cell fate control gene therapy

Cited by 8 publications

References 38 publications

Autologous, lentivirus‐modified, T‐rapa cell “micropharmacies” for lysosomal storage disorders

Autologous, lentivirus‐modified, T‐rapa cell “micropharmacies” for lysosomal storage disorders

S14G‐humanin alleviates insulin resistance and increases autophagy in neurons of APP/PS1 transgenic mouse

Rapamycin‐resistant effector T‐cell therapy

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