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Brain-Derived Extracellular Vesicles as Therapeutic Vehicles and Molecular Probes in the Neonatal Brain
Brain-Derived Extracellular Vesicles as Therapeutic Vehicles and Molecular Probes in the Neonatal Brain
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211152039
- ISBN
- 9798384094104
- DDC
- 610
- 서명/저자
- Brain-Derived Extracellular Vesicles as Therapeutic Vehicles and Molecular Probes in the Neonatal Brain
- 발행사항
- [Sl] : University of Washington, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 151 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 86-03, Section: B.
- 주기사항
- Advisor: Nance, Elizabeth.
- 학위논문주기
- Thesis (Ph.D.)--University of Washington, 2024.
- 초록/해제
- 요약In the central nervous system (CNS), intercellular communication through extracellular vesicles (EVs) is crucial for sustained trauma response and tissue repair following injury. EVs are biologically derived nanoparticles released by every cell that carry a diverse cargo of biomolecules important for cell communication including proteins, lipids, carbohydrates, and genetic material. The contents of EV cargo is an active research question in the field, and has been shown to be dependent on cell type, as well as environmental and physiological changes. As EVs are both produced and trafficked by cells, they are strong therapeutic candidates with several inherent design advantages over existing nanoparticle therapeutics: biostability, biocompatibility, lipid bilayer protection of cargo, and inherent cell uptake mechanisms. These advantages are especially important for drug delivery to the brain, which presents several therapeutic barriers such as the cerebral spinal fluid barrier and tortuous brain parenchyma. In addition, the blood brain barrier (BBB) poses a great challenge for therapeutics researchers as it excludes 98% of all small molecule and macromolecular drugs from passing but EVs have demonstrated an ability to cross. Despite growing interests in advancing EV therapeutics for brain injury and disease, there are two significant challenges hindering their development and clinical translation: 1) lack of physiologically relevant EV models and 2) a need for greater clarity about EV localization and transport in brain tissue. To address the first challenge, I evaluated the therapeutic efficacy of EVs derived from brain tissue (BEVs), rather than the standard approach of using EVs from cell culture. Compared to EVs derived from 2D cell monoculture models, those derived from 3D tissue are more physiologically relevant as they represent a heterogenous population that mirrors the existence of diverse cell types found in native brain tissue. When evaluating the therapeutic potential of BEVs in an ex vivo model of oxygen glucose deprivation (OGD), BEVs exhibited dose- and time-dependent therapeutic effects on injured tissue. BEVs induced a shift in the microglial morphology of OGD tissues from an inflammatory towards a restorative phenotype, while simultaneously increasing anti-inflammatory cytokine expression and decreasing cell cytotoxicity. These promising results led to further studies to address the second challenge-a lack in understanding about BEV localization and transport.To track BEVs in the brain we conjugated BEVs to either quantum dots (QDs) or novel oligonucleotide biobarcodes (oligobarcodes) using an efficient click chemistry reaction. Through a combination of confocal imaging and multiple particle tracking, the QD conjugation allowed us to visualize the spatial distribution of BEVs in brain tissue, which was regionally dependent. QD conjugations also allowed us to track BEV transport properties in real-time to confirm that BEV behavior was regionally dependent. We then used oligobarcoded BEVs to quantitatively measure the BEV uptake in both glial and non-glial cells in healthy and OGD brain tissue. Microglia, the resident immune cell of the brain, exhibited increased and preferential uptake of oligobarcoded BEVs compared to blank oligobarcode controls. We then expanded our oligobarcode-EV conjugation strategy to study the uptake of semen-derived EVs in the vaginal tract, demonstrating the broad translational opportunities that our platform provided to tracking EVs from any source. Collectively this work demonstrates the therapeutic potential of tissue-derived BEVs and offers a dual-conjugation technique to visualize and track EVs from any source in physiological environments. Our novel QD and oligobarcode conjugation strategy is an accessible technique that can be translated across different biological models to provide both quantitative and qualitative evaluation of EV visualization and tracking that will advance the EV therapeutics landscape.
- 일반주제명
- Bioengineering
- 일반주제명
- Nanoscience
- 일반주제명
- Neurosciences
- 일반주제명
- Genetics
- 키워드
- Microglia
- 키워드
- Quantum dot
- 기타저자
- University of Washington Molecular Engineering and Sciences
- 기본자료저록
- Dissertations Abstracts International. 86-03B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520250211152039
■006m o d
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■020 ▼a9798384094104
■035 ▼a(MiAaPQ)AAI31336104
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a610
■1001 ▼aNguyen, Nam Phuong.
■24510▼aBrain-Derived Extracellular Vesicles as Therapeutic Vehicles and Molecular Probes in the Neonatal Brain
■260 ▼a[Sl]▼bUniversity of Washington▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a151 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 86-03, Section: B.
■500 ▼aAdvisor: Nance, Elizabeth.
■5021 ▼aThesis (Ph.D.)--University of Washington, 2024.
■520 ▼aIn the central nervous system (CNS), intercellular communication through extracellular vesicles (EVs) is crucial for sustained trauma response and tissue repair following injury. EVs are biologically derived nanoparticles released by every cell that carry a diverse cargo of biomolecules important for cell communication including proteins, lipids, carbohydrates, and genetic material. The contents of EV cargo is an active research question in the field, and has been shown to be dependent on cell type, as well as environmental and physiological changes. As EVs are both produced and trafficked by cells, they are strong therapeutic candidates with several inherent design advantages over existing nanoparticle therapeutics: biostability, biocompatibility, lipid bilayer protection of cargo, and inherent cell uptake mechanisms. These advantages are especially important for drug delivery to the brain, which presents several therapeutic barriers such as the cerebral spinal fluid barrier and tortuous brain parenchyma. In addition, the blood brain barrier (BBB) poses a great challenge for therapeutics researchers as it excludes 98% of all small molecule and macromolecular drugs from passing but EVs have demonstrated an ability to cross. Despite growing interests in advancing EV therapeutics for brain injury and disease, there are two significant challenges hindering their development and clinical translation: 1) lack of physiologically relevant EV models and 2) a need for greater clarity about EV localization and transport in brain tissue. To address the first challenge, I evaluated the therapeutic efficacy of EVs derived from brain tissue (BEVs), rather than the standard approach of using EVs from cell culture. Compared to EVs derived from 2D cell monoculture models, those derived from 3D tissue are more physiologically relevant as they represent a heterogenous population that mirrors the existence of diverse cell types found in native brain tissue. When evaluating the therapeutic potential of BEVs in an ex vivo model of oxygen glucose deprivation (OGD), BEVs exhibited dose- and time-dependent therapeutic effects on injured tissue. BEVs induced a shift in the microglial morphology of OGD tissues from an inflammatory towards a restorative phenotype, while simultaneously increasing anti-inflammatory cytokine expression and decreasing cell cytotoxicity. These promising results led to further studies to address the second challenge-a lack in understanding about BEV localization and transport.To track BEVs in the brain we conjugated BEVs to either quantum dots (QDs) or novel oligonucleotide biobarcodes (oligobarcodes) using an efficient click chemistry reaction. Through a combination of confocal imaging and multiple particle tracking, the QD conjugation allowed us to visualize the spatial distribution of BEVs in brain tissue, which was regionally dependent. QD conjugations also allowed us to track BEV transport properties in real-time to confirm that BEV behavior was regionally dependent. We then used oligobarcoded BEVs to quantitatively measure the BEV uptake in both glial and non-glial cells in healthy and OGD brain tissue. Microglia, the resident immune cell of the brain, exhibited increased and preferential uptake of oligobarcoded BEVs compared to blank oligobarcode controls. We then expanded our oligobarcode-EV conjugation strategy to study the uptake of semen-derived EVs in the vaginal tract, demonstrating the broad translational opportunities that our platform provided to tracking EVs from any source. Collectively this work demonstrates the therapeutic potential of tissue-derived BEVs and offers a dual-conjugation technique to visualize and track EVs from any source in physiological environments. Our novel QD and oligobarcode conjugation strategy is an accessible technique that can be translated across different biological models to provide both quantitative and qualitative evaluation of EV visualization and tracking that will advance the EV therapeutics landscape.
■590 ▼aSchool code: 0250.
■650 4▼aBioengineering
■650 4▼aNanoscience
■650 4▼aNeurosciences
■650 4▼aGenetics
■653 ▼aExtracellular vesicles
■653 ▼aMicroglia
■653 ▼aMultiple particle tracking
■653 ▼aNeonatal hypoxia ischemia
■653 ▼aOligonucleotide barcode
■653 ▼aQuantum dot
■690 ▼a0202
■690 ▼a0565
■690 ▼a0317
■690 ▼a0369
■71020▼aUniversity of Washington▼bMolecular Engineering and Sciences.
■7730 ▼tDissertations Abstracts International▼g86-03B.
■790 ▼a0250
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17162664▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
