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HyeongJin Yun Scientific Essay

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'''Necessity of RNA Sequencing in Human BBB Models''' '''&~~> and Drug Delivery Research''' 20191193 HyeongJin Yun '''Introduction''' I have always been deeply interested in diseases caused by brain dysfunction,~~sans-serif"="">Introduction''' <span style="font~~such as brain cancer, depression, and dementia. This interest led me to pursue research and development in the field of drug delivery to the brain, particularly through the blood-~~size:10pt">I am currently developing a human blood brain barrier (BBB) model~~ brain barrier (BBB) model and conducting extensive research to understand how carriers penetrate the BBB for drug delivery. My work primarily involves building basic cell culture techniques and creating an in vitro BBB model. The carrier samples used in this experiment are provided by our collaborative research partner. We plan and ~~conducting extensive research to understand how~~ execute further experimental procedures by determining the permeability of these carriers ~~penetrate~~ across the BBB ~~for drug delivery~~. My work is primarily to build basic cell culture techniques and an in vitro BBB model. The carrier samples used in this experiment are provided by our collaborative research partner. We plan and execute further experimental procedures by determining Currently, I have completed the ~~permeability~~ process of ~~these carriers across~~ determining whether lipid nanoparticles (LNPs) can pass through the BBB.~~ <span style="font~~Now, I have shifted my focus to exploring the mechanisms by which these nanoparticles pass through the barrier. I became particularly interested in this area because I believe RNA-~~size:10pt"><span~~ sequencing can provide detailed insights into the gene expression changes and molecular pathways activated during nanoparticle transport across the BBB, thereby elucidating the specific biological processes and interactions involved. '''Blood-~~family:" arial~~Brain Barrier and Lipid Nanoparticles''' ~~Currently, I have completed the process of determining whether lipid nanoparticles~~ The human blood-brain barrier (~~LNPs~~BBB) ~~can pass through BBB. Now I have shifted my focus~~ is a unique and selective barrier that regulates the transport of substances from Blood to ~~exploring~~ the ~~mechanisms by which these nanoparticles pass through~~ brain. It plays an important role in maintaining the ~~barrier~~neurons and glia function. ~~I came to find out because I thought RNA-sequencing~~BBB is composed of Brain microvascular endothermic cells (BMVECs), Astrocytes, ~~which was mentioned in Genomics course~~Pericytes, ~~would help me explore this mechanism.~~and ECM (</spanstyle="color:#0070c0">1,2~~<~~). BMVECs have a much lower degree of endocytosis/transcytosis activity than peripheral endothelium contributing to the characteristics of BBB as a barrier (<spanstyle="color:#0070c0">3<br/span> ).<br/span> ''' Lipid nanoparticles (LNPs) are widely used in the field of drug delivery. LNPs are primarily composed of cholesterol and lipids and can encapsulate DNA,~~sans-serif"~~RNA, or drugs. Additionally, ligands or antibodies can be attached to the surface to aid in targeting (~~Blood-Brain Barrier and Lipid Nanoparticles'''~~4). ~~The~~ Assembly (Luca) can pass through the human ~~blood-brain barrier (~~BBB) model. The identification of LNP samples that penetrate well is ~~a unique~~ finished and ~~selective barrier that regulates~~ the ~~transport of substances from Blood to~~ mechanism study process is in the ~~brain~~process. ~~It plays an important role in maintaining the neurons and glia function. BBB is composed of Brain microvascular endothermic cells (BMVECs), Astrocytes, Pericytes, and ECM (~~ '''~~1,2~~Overview of RNA sequencing technology~~). BMVECs have a much lower degree of endocytosis~~'''</~~transcytosis activity than peripheral endothelium contributing to the characteristics of BBB as a barrier (~~span><~~span style~~br/> 3<~~/span>).</~~spanlang="EN-US">Principles of RNA Sequencing<br/~~> ~~ Lipid nanoparticles (LNPs) are widely~~ throughput sequencing technology used in to determine the ~~field~~ nucleotide sequence of ~~drug delivery~~RNA molecules and quantify specific RNA species within a population. ~~LNPs are primarily composed~~ The process of ~~cholesterol and lipids and can encapsulate DNA,~~ RNA~~, or drugs. Additionally, ligands or antibodies can be attached to~~ sequencing includes the ~~surface to aid in targeting~~ following steps (45).:~~ ~~ #RNA Extraction: Extracting RNA from biological samples such as cells or tissues.  <ol start="2"><li>My research is conducting a study on whether LNPs using a new fabrication method called Liposome Under Cryo-Assembly (Luca) can pass through the human BBB model. The identification of LNP samples that penetrate well is finished and the mechanism study process is in the process. '''Overview of RNA sequencing technology''' Principles of RNA Sequencing RNA sequencing (RNA-seq) is a high-throughput sequencing technology used to determine the nucleotide sequence of RNA molecules and quantify specific RNA species within a population. The process of RNA sequencing includes the following steps (5): ~~#RNA Extraction~~size: ~~Extracting RNA from biological samples such as cells or tissues.~~ ~~&nbsp~~14px; ~~<ol start="2"><li>cDNA Synthesis: Converting the extracted RNA into complementary DNA (cDNA) using reverse transcriptase.~~</li>

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~~ ~~  <ol start="3"><li>Library Preparation: Fragmenting the cDNA, attaching adapters, and amplifying the cDNA fragments through PCR.~~~~</li>

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<ol start="4"><li>Sequencing: Sequencing the prepared library using platforms such as Illumina or Nanopore.~~~~</li>

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<ol start="5"><li>Data Analysis: Aligning the generated sequences to a reference genome and quantifying gene expression levels.~~~~</li>

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'''Methods for Gene Expression Analysis Using RNA Sequencing'''~~~~<br/~~> The analysis of gene expression using RNA sequencing involves several steps:~~~~   #Data Quality Control: Assessing the quality of sequencing data and removing low-quality reads.~~~~   <ol start="2"><li>Data Alignment: Aligning the reads to a reference genome to determine the location of each read.~~</li>

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~~ ~~  <ol start="3"><li>Gene Expression Quantification: Calculating the expression levels of genes, typically represented as RPKM, FPKM, or TPM values.~~~~</li>

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<ol start="4"><li>Differential Expression Analysis: Identifying differentially expressed genes between specific conditions to understand changes in gene expression due to factors such as inflammation, stress, or drug treatment~~~~</li>

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'''Necessity of RNA Sequencing in BBB Models'''~~~~<br/~~> Through fluorescence image analysis, I found out that LNPs migrate to the transcytosis pathway. In addition, I checked the difference in transmittance by performing several transcytosis inhibitor treatments.~~~~<br/~~> Through RNA sequencing, I would like to check whether the permeability has changed due to changes in the genetic level of the cells that actually make up BBB.~~~~ <br/~~> '''Future Plan'''~~~~<br/~~> Listening to the Genomics course, I thought that conducting research at various gene levels was essential to conduct research at a high level. Later, the goal is to proceed with the RNA sequencing mentioned above to find out the detailed mechanisms that have not yet been found. After finding out the mechanism, I will treat the Aptamer on the surface to find out the change in the permeability and put the actual drug inside to deliver it.~~~~ <br/~~> '''References'''~~~~<br/~~> (1) Cecchelli, Romeo, et al., Modelling of the blood–brain barrier in drug discovery and development., '''2007''', ''Nature Reviews Drug Discovery''. vol. 6, no. 8, pp. 650–661,.~~~~ <br/~~> (2) Park, Tae-Eun, et al., Hypoxia-enhanced blood-brain barrier chip recapitulates human barrier function and shuttling of drugs and antibodies., '''2019''', ''Nature Communications'', vol. 10, no. 1, 13~~~~ <br/~~> (3) Abbott, N. Joan, Lars Rönnbäck, and Elisabeth Hansson., Astrocyte–Endothelial Interactions at the Blood–Brain Barrier., '''2006''', ''Nature Reviews Neuroscience 7'', no. 1: 41–53.~~~~ <br/~~> (4) Mehta, M., Bui, T. A., Yang, X., Aksoy, Y., Goldys, E. M., & Deng, W., Lipid-based nanoparticles for drug/gene delivery: An overview of the production techniques and difficulties encountered in their industrial development., '''2023''', ''ACS Materials Au'', 3(6), 600–619.~~~~ <br/~~> (5) Deshpande, Dhrithi, et al., RNA-seq data science: From RAW data to effective interpretation.,'''2023''', ''Frontiers in Genetics'', vol. 14,~~~~

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