Multiplex Genome Modifications of Astrocytes Through CRISPR Dead Cas9: A Novel Candidate Therapy for Chronic Ischemic Stroke
Abstract
A stroke is an enervating injury to the brain that occurs from a stoppage in blood supply (ischemic stroke) or bleeding (hemorrhagic) in a hemisphere of the brain. Globally, about 10 million deaths per year are recorded because of stroke. There has been no definitive FDA-approved treatment for chronic ischemic stroke without any side effects so far. Therefore, the search for new therapies is necessary. In this paper, after investigating several studies online, on Google Scholar, PubMed, and Scopus, we hypothesized improving the complications of chronic ischemic stroke in induced Sprague-Dawley rat model by intraluminal suture middle cerebral artery occlusion (MCAo), utilizing the combination of cell therapy and gene therapy.
A new version of astrocytes is proposed by making some changes in their genome. To gain this goal, a gene profile including IL-38 (the most modern anti-inflammatory agent, which barricades inflammatory response factors), BRAG-1 (an anti-apoptotic gene from BCL-2 family), IL-38 and BRAG-1’s complementary scaffold RNAs for their expression by deadCas9 (dCas9), complementary scaffold RNAs of LZK and MST-1 for their deletion, and deadCas9 gene is used.
Based on studies and documents, we hypothesized using modified astrocytes by dCas9, which is the most accurate genome-editing technology
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3. Li M, Chen S, Shi X, Lyu C, Zhang Y, Tan M, et al. Cell permeable HMGB1-binding heptamer peptide ameliorates neurovascular complications associated with thrombolytic therapy in rats with transient ischemic stroke. Journal of neuroinflammation. 2018;15(1):237.
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15. Buffo A, Rite I, Tripathi P, Lepier A, Colak D, Horn A-P, et al. Origin and progeny of reactive gliosis: A source of multipotent cells in the injured brain. Proceedings of the National Academy of Sciences. 2008;105(9):3581-6.
16. Duan CL, Liu CW, Shen SW, Yu Z, Mo JL, Chen XH, et al. Striatal astrocytes transdifferentiate into functional mature neurons following ischemic brain injury. Glia. 2015;63(9):1660-70.
17. Verkhratsky A, Untiet V, Rose CR. Ionic signalling in astroglia beyond calcium. The Journal of physiology. 2019.
18. Walz W. Role of astrocytes in the clearance of excess extracellular potassium. Neurochemistry international. 2000;36(4-5):291-300.
19. D'Ambrosio R, Gordon DS, Winn HR. Differential role of KIR channel and Na+/K+-pump in the regulation of extracellular K+ in rat hippocampus. Journal of neurophysiology. 2002;87(1):87-102.
20. Abbott NJ, Rönnbäck L, Hansson E. Astrocyte–endothelial interactions at the blood–brain barrier. Nature reviews neuroscience. 2006;7(1):41.
21. Campisi M, Shin Y, Osaki T, Hajal C, Chiono V, Kamm RD. 3D BLOOD-BRAIN BARRIER MICROVASCULAR NETWORK MODEL INCLUDING HUMAN IPS-DERIVED ENDOTHELIAL CELLS, PERICYTES AND ASTROCYTES. 2019.
22. Newman EA. Glial cell regulation of neuronal activity and blood flow in the retina by release of gliotransmitters. Philosophical Transactions of the Royal Society B: Biological Sciences. 2015;370(1672):20140195.
23. Zeng X-N, Sun X-L, Gao L, Fan Y, Ding J-H, Hu G. Aquaporin-4 deficiency down-regulates glutamate uptake and GLT-1 expression in astrocytes. Molecular and Cellular Neuroscience. 2007;34(1):34-9.
24. Kitchen P, Day RE, Taylor LH, Salman MM, Bill RM, Conner MT, et al. Identification and molecular mechanisms of the rapid tonicity-induced relocalization of the aquaporin 4 channel. Journal of Biological Chemistry. 2015;290(27):16873-81.
25. Pentreath V, Slamon N. Astrocyte phenotype and prevention against oxidative damage in neurotoxicity. Human & experimental toxicology. 2000;19(11):641-9.
26. Dong Y, Benveniste EN. Immune function of astrocytes. Glia. 2001;36(2):180-90.
27. Attwell D, Buchan AM, Charpak S, Lauritzen M, MacVicar BA, Newman EA. Glial and neuronal control of brain blood flow. Nature. 2010;468(7321):232.
28. Halassa MM, Haydon PG. Integrated brain circuits: astrocytic networks modulate neuronal activity and behavior. Annual review of physiology. 2010;72:335-55.
29. Verkhratsky A, Orkand RK, Kettenmann H. Glial calcium: homeostasis and signaling function. Physiological reviews. 1998;78(1):99-141.
30. Verkhratsky A. Astroglial Calcium Signaling in Aging and Alzheimer's Disease. Cold Spring Harbor Perspectives in Biology. 2019:a035188.
31. Pekny M, Nilsson M. Astrocyte activation and reactive gliosis. Glia. 2005;50(4):427-34.
32. Robel S, Berninger B, Götz M. The stem cell potential of glia: lessons from reactive gliosis. Nature Reviews Neuroscience. 2011;12(2):88.
33. Chen M, Geoffroy CG, Meves JM, Narang A, Li Y, Nguyen MT, et al. Leucine zipper-bearing kinase is a critical regulator of astrocyte reactivity in the adult mammalian CNS. Cell reports. 2018;22(13):3587-97.
34. Briens A, Bardou I, Lebas H, Miles LA, Parmer RJ, Vivien D, et al. Astrocytes regulate the balance between plasminogen activation and plasmin clearance via cell-surface actin. Cell discovery. 2017;3:17001.
35. Zalatan JG, Lee ME, Almeida R, Gilbert LA, Whitehead EH, La Russa M, et al. Engineering complex synthetic transcriptional programs with CRISPR RNA scaffolds. 2015;160(1-2):339-50.
36. Ali Z, Mahas A, Mahfouz M. CRISPR/Cas13 as a tool for RNA interference. Trends in plant science. 2018;23(5):374-8.
37. Kim H, Kim J-S. A guide to genome engineering with programmable nucleases. Nature Reviews Genetics. 2014;15(5):321-34.
38. Pawluk A, Amrani N, Zhang Y, Garcia B, Hidalgo-Reyes Y, Lee J, et al. Naturally occurring off-switches for CRISPR-Cas9. Cell. 2016;167(7):1829-38. e9.
39. Fluri F, Schuhmann MK, Kleinschnitz C. Animal models of ischemic stroke and their application in clinical research. Drug design, development and therapy. 2015;9:3445.
40. Yuan X, Peng X, Li Y, Li M. Role of IL-38 and its related cytokines in inflammation. Mediators of inflammation. 2015;2015.
41. Esmaeilzadeh A, Pouyan S, Erfanmanesh M. Is Interleukin-38 a key player cytokine in atherosclerosis immune gene therapy? Medical hypotheses. 2019;125:139-43.
42. Scherz P. The mechanism and applications of CRISPR-Cas9. The National Catholic Bioethics Quarterly. 2017;17(1):29-36.
43. Nakamura-Ishizu A, Kurihara T, Okuno Y, Ozawa Y, Kishi K, Goda N, et al. The formation of an angiogenic astrocyte template is regulated by the neuroretina in a HIF-1-dependent manner. Developmental biology. 2012;363(1):106-14.
44. Synthego. CRISPR 101 Your Guide to Understanding CRISPR 2019.
45. Mohamed H. The Effect of Buffers Ph on Electrophoretic Purification of Intracellular Green Fluorescent Protein: UMP; 2014.
46. Mecha M, Iñigo PM, Mestre L, Hernangómez M, Borrell J, Guaza C. An easy and fast way to obtain a high number of glial cells from rat cerebral tissue: a beginners approach. Protocol Exchange. 2011;218:1038.
47. Cao L-M, Dong Z-Q, Li Q, Chen X. Treadmill training improves neurological deficits and suppresses neuronal apoptosis in cerebral ischemic stroke rats. Neural regeneration research. 2019;14(8):1387.
48. Haas S, Weidner N, Winkler J. Adult stem cell therapy in stroke. Current opinion in neurology. 2005;18(1):59-64.
49. Gao J, Dennis JE, Muzic RF, Lundberg M, Caplan AI. The dynamic in vivo distribution of bone marrow-derived mesenchymal stem cells after infusion. Cells Tissues Organs. 2001;169(1):12-20.
50. Tsuji O, Miura K, Okada Y, Fujiyoshi K, Mukaino M, Nagoshi N, et al. Therapeutic potential of appropriately evaluated safe-induced pluripotent stem cells for spinal cord injury. Proceedings of the National Academy of Sciences. 2010;107(28):12704-9.
51. Tang Q, Chen Q, Lai X, Liu S, Chen Y, Zheng Z, et al. Malignant transformation potentials of human umbilical cord mesenchymal stem cells both spontaneously and via 3-methycholanthrene induction. PloS one. 2013;8(12).
2. Casals JB, Pieri NC, Feitosa ML, Ercolin A, Roballo K, Barreto RS, et al. The use of animal models for stroke research: a review. Comparative medicine. 2011;61(4):305-13.
3. Li M, Chen S, Shi X, Lyu C, Zhang Y, Tan M, et al. Cell permeable HMGB1-binding heptamer peptide ameliorates neurovascular complications associated with thrombolytic therapy in rats with transient ischemic stroke. Journal of neuroinflammation. 2018;15(1):237.
4. Leng T, Xiong Z-G. Treatment for ischemic stroke: From thrombolysis to thrombectomy and remaining challenges. Brain Circulation. 2019;5(1):8.
5. Deb P, Sharma S, Hassan K. Pathophysiologic mechanisms of acute ischemic stroke: An overview with emphasis on therapeutic significance beyond thrombolysis. Pathophysiology. 2010;17(3):197-218.
6. Sarvari S, Moakedi F, Hone E, Simpkins JW, Ren X. Mechanisms in blood-brain barrier opening and metabolism-challenged cerebrovascular ischemia with emphasis on ischemic stroke. Metabolic brain disease. 2020.
7. Nozohouri S, Sifat AE, Vaidya B, Abbruscato TJ. Novel approaches for the delivery of therapeutics in ischemic stroke. Drug Discovery Today. 2020.
8. Ooboshi H. Gene therapy as a novel pharmaceutical intervention for stroke. Current pharmaceutical design. 2011;17(5):424-33.
9. Marei HE, Hasan A, Rizzi R, Althani A, Afifi N, Cenciarelli C, et al. Potential of stem cell-based therapy for ischemic stroke. Frontiers in neurology. 2018;9:34.
10. Boese AC, Le Q-SE, Pham D, Hamblin MH, Lee J-P. Neural stem cell therapy for subacute and chronic ischemic stroke. Stem cell research & therapy. 2018;9(1):154.
11. Danbolt NC. Glutamate uptake. Progress in neurobiology. 2001;65(1):1-105.
12. Da Silva APB, de Souza DG, Souza DO, Machado DC, Sato DK. Role of Glutamatergic Excitotoxicity in Neuromyelitis Optica Spectrum Disorders. Frontiers in cellular neuroscience. 2019;13:142.
13. Schneider J, Karpf J, Beckervordersandforth R. Role of Astrocytes in the Neurogenic Niches. Astrocytes: Springer; 2019. p. 19-33.
14. Mori T, Buffo A, Götz M. The novel roles of glial cells revisited: the contribution of radial glia and astrocytes to neurogenesis. Current topics in developmental biology. 2005;69:67-99.
15. Buffo A, Rite I, Tripathi P, Lepier A, Colak D, Horn A-P, et al. Origin and progeny of reactive gliosis: A source of multipotent cells in the injured brain. Proceedings of the National Academy of Sciences. 2008;105(9):3581-6.
16. Duan CL, Liu CW, Shen SW, Yu Z, Mo JL, Chen XH, et al. Striatal astrocytes transdifferentiate into functional mature neurons following ischemic brain injury. Glia. 2015;63(9):1660-70.
17. Verkhratsky A, Untiet V, Rose CR. Ionic signalling in astroglia beyond calcium. The Journal of physiology. 2019.
18. Walz W. Role of astrocytes in the clearance of excess extracellular potassium. Neurochemistry international. 2000;36(4-5):291-300.
19. D'Ambrosio R, Gordon DS, Winn HR. Differential role of KIR channel and Na+/K+-pump in the regulation of extracellular K+ in rat hippocampus. Journal of neurophysiology. 2002;87(1):87-102.
20. Abbott NJ, Rönnbäck L, Hansson E. Astrocyte–endothelial interactions at the blood–brain barrier. Nature reviews neuroscience. 2006;7(1):41.
21. Campisi M, Shin Y, Osaki T, Hajal C, Chiono V, Kamm RD. 3D BLOOD-BRAIN BARRIER MICROVASCULAR NETWORK MODEL INCLUDING HUMAN IPS-DERIVED ENDOTHELIAL CELLS, PERICYTES AND ASTROCYTES. 2019.
22. Newman EA. Glial cell regulation of neuronal activity and blood flow in the retina by release of gliotransmitters. Philosophical Transactions of the Royal Society B: Biological Sciences. 2015;370(1672):20140195.
23. Zeng X-N, Sun X-L, Gao L, Fan Y, Ding J-H, Hu G. Aquaporin-4 deficiency down-regulates glutamate uptake and GLT-1 expression in astrocytes. Molecular and Cellular Neuroscience. 2007;34(1):34-9.
24. Kitchen P, Day RE, Taylor LH, Salman MM, Bill RM, Conner MT, et al. Identification and molecular mechanisms of the rapid tonicity-induced relocalization of the aquaporin 4 channel. Journal of Biological Chemistry. 2015;290(27):16873-81.
25. Pentreath V, Slamon N. Astrocyte phenotype and prevention against oxidative damage in neurotoxicity. Human & experimental toxicology. 2000;19(11):641-9.
26. Dong Y, Benveniste EN. Immune function of astrocytes. Glia. 2001;36(2):180-90.
27. Attwell D, Buchan AM, Charpak S, Lauritzen M, MacVicar BA, Newman EA. Glial and neuronal control of brain blood flow. Nature. 2010;468(7321):232.
28. Halassa MM, Haydon PG. Integrated brain circuits: astrocytic networks modulate neuronal activity and behavior. Annual review of physiology. 2010;72:335-55.
29. Verkhratsky A, Orkand RK, Kettenmann H. Glial calcium: homeostasis and signaling function. Physiological reviews. 1998;78(1):99-141.
30. Verkhratsky A. Astroglial Calcium Signaling in Aging and Alzheimer's Disease. Cold Spring Harbor Perspectives in Biology. 2019:a035188.
31. Pekny M, Nilsson M. Astrocyte activation and reactive gliosis. Glia. 2005;50(4):427-34.
32. Robel S, Berninger B, Götz M. The stem cell potential of glia: lessons from reactive gliosis. Nature Reviews Neuroscience. 2011;12(2):88.
33. Chen M, Geoffroy CG, Meves JM, Narang A, Li Y, Nguyen MT, et al. Leucine zipper-bearing kinase is a critical regulator of astrocyte reactivity in the adult mammalian CNS. Cell reports. 2018;22(13):3587-97.
34. Briens A, Bardou I, Lebas H, Miles LA, Parmer RJ, Vivien D, et al. Astrocytes regulate the balance between plasminogen activation and plasmin clearance via cell-surface actin. Cell discovery. 2017;3:17001.
35. Zalatan JG, Lee ME, Almeida R, Gilbert LA, Whitehead EH, La Russa M, et al. Engineering complex synthetic transcriptional programs with CRISPR RNA scaffolds. 2015;160(1-2):339-50.
36. Ali Z, Mahas A, Mahfouz M. CRISPR/Cas13 as a tool for RNA interference. Trends in plant science. 2018;23(5):374-8.
37. Kim H, Kim J-S. A guide to genome engineering with programmable nucleases. Nature Reviews Genetics. 2014;15(5):321-34.
38. Pawluk A, Amrani N, Zhang Y, Garcia B, Hidalgo-Reyes Y, Lee J, et al. Naturally occurring off-switches for CRISPR-Cas9. Cell. 2016;167(7):1829-38. e9.
39. Fluri F, Schuhmann MK, Kleinschnitz C. Animal models of ischemic stroke and their application in clinical research. Drug design, development and therapy. 2015;9:3445.
40. Yuan X, Peng X, Li Y, Li M. Role of IL-38 and its related cytokines in inflammation. Mediators of inflammation. 2015;2015.
41. Esmaeilzadeh A, Pouyan S, Erfanmanesh M. Is Interleukin-38 a key player cytokine in atherosclerosis immune gene therapy? Medical hypotheses. 2019;125:139-43.
42. Scherz P. The mechanism and applications of CRISPR-Cas9. The National Catholic Bioethics Quarterly. 2017;17(1):29-36.
43. Nakamura-Ishizu A, Kurihara T, Okuno Y, Ozawa Y, Kishi K, Goda N, et al. The formation of an angiogenic astrocyte template is regulated by the neuroretina in a HIF-1-dependent manner. Developmental biology. 2012;363(1):106-14.
44. Synthego. CRISPR 101 Your Guide to Understanding CRISPR 2019.
45. Mohamed H. The Effect of Buffers Ph on Electrophoretic Purification of Intracellular Green Fluorescent Protein: UMP; 2014.
46. Mecha M, Iñigo PM, Mestre L, Hernangómez M, Borrell J, Guaza C. An easy and fast way to obtain a high number of glial cells from rat cerebral tissue: a beginners approach. Protocol Exchange. 2011;218:1038.
47. Cao L-M, Dong Z-Q, Li Q, Chen X. Treadmill training improves neurological deficits and suppresses neuronal apoptosis in cerebral ischemic stroke rats. Neural regeneration research. 2019;14(8):1387.
48. Haas S, Weidner N, Winkler J. Adult stem cell therapy in stroke. Current opinion in neurology. 2005;18(1):59-64.
49. Gao J, Dennis JE, Muzic RF, Lundberg M, Caplan AI. The dynamic in vivo distribution of bone marrow-derived mesenchymal stem cells after infusion. Cells Tissues Organs. 2001;169(1):12-20.
50. Tsuji O, Miura K, Okada Y, Fujiyoshi K, Mukaino M, Nagoshi N, et al. Therapeutic potential of appropriately evaluated safe-induced pluripotent stem cells for spinal cord injury. Proceedings of the National Academy of Sciences. 2010;107(28):12704-9.
51. Tang Q, Chen Q, Lai X, Liu S, Chen Y, Zheng Z, et al. Malignant transformation potentials of human umbilical cord mesenchymal stem cells both spontaneously and via 3-methycholanthrene induction. PloS one. 2013;8(12).
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| Issue | Vol 7, No 4 (2024) | |
| Section | Review Article | |
| DOI | https://doi.org/10.18502/igj.v7i4.17890 | |
| Keywords | ||
| Astrocytes CRISPR Cell Therapy Chronic Ischemic Stroke Gene Therapy Scaffold Rnas | ||
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How to Cite
1.
Lotfi M. Multiplex Genome Modifications of Astrocytes Through CRISPR Dead Cas9: A Novel Candidate Therapy for Chronic Ischemic Stroke. Immunol Genet J. 2024;7(4):260-267.
