4 research outputs found
νμλ§μΆ€ν μΉλ£λ₯Ό μν μ²΄μΈ νμμ μ€ν¬λ¦¬λμ© λ°μ΄μ€μΉ©
νμλ
Όλ¬Έ (λ°μ¬) -- μμΈλνκ΅ λνμ : 곡과λν μ κΈ°Β·μ 보곡νλΆ, 2020. 8. κΆμ±ν.μ λ°μν(Precision Medicine) νΉμ κ°μΈλ§μΆ€μν(Personalized Medicine)μ κ°κ°μΈμ μ΅μ νλ μΉλ£λ°©λ²μ κ²°μ νλ κ²μ λͺ©νλ‘ νλ μνμ ν¨λ¬λ€μμ΄λ€. νΉν, μμμ’
μνμμλ μ°¨μΈλμΌκΈ°μμ΄λΆμ(NGS), μ μ¬μ²΄μμ΄λΆμ, κ·Έλ¦¬κ³ μ§λλΆμλ²λ€μ ν΅ν νμμ λΆμ νλ‘νμΌ(molecular profile) λ°©λ²μ΄ λ°μ ν΄μ€κ³ μμΌλ©°, μ΄λ₯Ό λ°νμΌλ‘ νμλ₯Ό μΈλΆννμ¬ λ§μΆ€ν μΉλ£λ₯Ό ꡬννλ €κ³ λ
Έλ ₯ν΄μ€κ³ μλ€. νμ§λ§, μ¬μ ν ν μμ€μμ μ΄ν΄λμ§ λͺ»νλ μμ€μ μ’
μ μ΄μ§μ±(tumor heterogeneity)κ³Ό μ€λ μ²λ°©κΈ°λ‘μ κ°μ§ νμκ΅°λ€μ νμμ νλλ΄μ±(acquired resistance) λ±μ μμΈμΌλ‘ λ§μΆ€ν νμ μ²λ°©μ μ½μ§ μμ κ²½μ°κ° λ§λ€. μ΄λ¬ν κ²½μ° νμλ‘λΆν° μ»μ΄μ§ μμΈν¬, μ‘°μ§μΌλ‘λΆν° μ»μ΄μ§ μΌμ°¨μΈν¬ νΉμ μ²΄μΈ λ°°μλ μΈν¬, μ€νλ‘μ΄λ, μ₯κΈ°μ μ¬μ²΄ λ±μ μ΄μ©νμ¬ κ³ μλ€μ€μ½λ¬Όμ€ν¬λ¦¬λκΈ°μ μ ν΅ν λ§μΆ€ν νμμ λ₯Ό μ λ³ν΄λ΄λ μ²΄μΈ μ½λ¬Όμ§λ¨ κΈ°μ μ μκ°ν΄λΌ μ μλλ°, μ΄λ κΈ°μ‘΄μ μ μ 체 κΈ°λ°μ μλμ λ³νλμ΄ κ°κ°μ νμλ€μκ² λμ± μ ν©ν μΉλ£λ°©λ²μ μ°Ύλ κ²μ΄ κ°λ₯νκ² νλ€.
νμ§λ§ μ΄λ¬ν λͺ©μ μ κ³ μλ€μ€μ½λ¬Όμ€ν¬λ¦¬λκΈ°μ μ λμ νμ©κ°λ₯μ±μλ λΆκ΅¬νκ³ , κ΄λ²μν 보κΈκ³Ό νμ©μ΄ λκΈ°μλ μ μ½μ μ΄ λ§μλ€. κΈ°μ‘΄μ κ³ μλ€μ€μ½λ¬Όμ€ν¬λ¦¬λκΈ°μ μ λ§μ μμ μνμ΄ μλͺ¨λκ³ , κ°λΉμΌ μμ½μ μλͺ¨λλ μ μ§ μμλ€. κ²λ€κ°, μμ² κ°μ§ μ΄μμ μλ‘ λ€λ₯Έ λ¬Όμ§λ€μ νμνκΈ° μν΄ λ°λμ νμν κ³ κ°μ μλνλ μ‘체 μ΄λ°κΈ°(liquid handler) λ±μ΄ νμνμλλ°, μ΄λ¬ν λ¬Έμ λ‘ λν μ μ½μ¬, μ°κ΅¬μ λ±μ μ μΈνκ³ λ λμ
μ΄ μ½μ§κ° μμ κΈ°μ μ κ·Όμ±μ΄ μ νλμ΄ μμλ€.
λ³Έ μ°κ΅¬μμλ λ°λ체곡μ μμμ λ
Έκ΄κΈ°μ μ μ΄μ©νμ¬ κ°κ°μ μλ³ν μ μλ μ½λλ₯Ό κ°μ§κ³ μλ μ½λνλ νμ΄λλ‘μ € κΈ°λ°μ κ΄κ²½νμ±ν΄λ¦¬λ¨Έ λ―ΈμΈμ
μλ₯Ό λ§λ€μ΄, μ΄λ₯Ό μνλ μμΈν¬μ μ½λ¬Ό μ€ν¬λ¦¬λμ ν΄λ³΄κ³ μ νλ λ€μν μ½λ¬ΌλΌμ΄λΈλ¬λ¦¬λ₯Ό μ΄μ© κ°κ°μ μ½λνλ λ―ΈμΈμ
μμ ν‘μμμΌ μ½λ¬Ό-λ―ΈμΈμ
μ λΌμ΄λΈλ¬λ¦¬λ₯Ό μ μνλ€. κ·Έν, κ°λΉμΌ μ΄λ μ΄ μ μμ© μ€ν¬ν° νΉμ λμ€νμ μ₯λΉμμ΄ κ°λ¨ν μ기쑰립μ ν΅ν΄ λκ·λͺ¨μ λ€μν μ½λ¬Ό-νμ΄λλ‘μ € μ΄λ μ΄λ₯Ό μ μν μ μλ κΈ°μ μ κ°λ°νμλ€. λν, μλμ μΈν¬λ€ λ§μΌλ‘λ λ―ΈμΈμ°λ¬Ό(microwell) κΈ°λ°μ μΈν¬μΉ©μ λν¬νλ λ°©μμ κ°λ°νμμΌλ©°, μ΄λ₯Όν΅ν΄ μ½λ¬Ό-νμ΄λλ‘μ € μ΄λ μ΄μ λ―ΈμΈμ°λ¬ΌκΈ°λ°μ μΈν¬μΉ©μ κ²°ν©μΌλ‘ μλ°±-μμ²μ λ€μν μ΄μΈμ΄λ₯Ό μ μ μμ μνλ§μΌλ‘λ νλ²μ μνν μ μλ κ³ μλ€μ€μ½λ¬Όμ€ν¬λ¦¬λ κΈ°μ μ μνν μ μκ² λ§λ€μλ€.
λ³Έ μ°κ΅¬μμ μ μν μννλ μ²΄μΈ νμμ μ€ν¬λ¦¬λμ© μ½λ¬Όνλ«νΌμ λ€μκ³Ό κ°μ μμλ₯Ό κ°μ§λ€. μ μ μμ νμμΈν¬ νΉμ μνμ μμ μ μ©ν μ μλ, μ¬μ©νκΈ° μμ¬μ΄ κΈ°μ λ‘μ, κΈ°μ‘΄μ κ°λΉμΌ μ₯λΉ, μμ½μ μ¬μ©λμ νκΈ°μ μΌλ‘ μ€μΌ μ μλ κΈ°μ μ΄λ€. λ³Έ μ°κ΅¬μμ μ μλ κΈ°μ μ ν΅ν΄ κΈ°μ‘΄μ μ₯λΉλ₯Ό μ¬μ©ν λ μμ½μ κ°μ΄ λΉμΈκ±°λ, μ₯λΉμ κ°κ²©μ΄ λΉμΈμ, νΉμ λ€λ£¨κ³ μ νλ μνμ μμ΄ μ νμ μ΄μ΄μ κΈ°μ‘΄μ μ κ·ΌνκΈ° νλ€μλ λ€μν νμ μ°κ΅¬μ μ μ©ν μ μμΌλ©°, λ³μμμμ μμμ°κ΅¬ λ° μ€μ νμλ§μΆ€ν μΉλ£μ μ¬μ© λ μ μλ μ κ·Όμ±μ νκΈ°μ μΌλ‘ λμΌ μ μλ€. νΉν, λΉκ΅μ μ€,μ κ·λͺ¨μ μ°κ΅¬νκ²½μμλ λ€μν ν¬κ·ν νμμ λμΈν¬ νΉμ νμμ λμ€κ°λ
Έμ΄λ λ±κ³Ό μ λͺ©νμ¬ μ¬μ©λλ€λ©΄ λ³Έ νλ«νΌμ κ°λ₯μ±μ λμ± κ·Ήλν ν μ μμ κ²μΌλ‘ κΈ°λνλ€.Precision or Personalized Medicine is a medical paradigm aimed to determine optimal therapy for individual patient. In particular, clinical oncology has been using methods of molecular profiling for each patient through next-generation sequencing (NGS), mRNA-sequencing, and mass spectrometry, and has been trying to implement personalized treatment. However, personalized treatment based on molecular profiling to each patient is not always possible due to the high level of heterogeneity of tumor that is still not fully understood at the current level and acquired resistance of anti-cancer drug due to cumulative targeted therapy. In such cases, in vitro drug testing platform using primary cells obtained from patients, or patient-derived cells, spheroids, and organoids can make it possible to find a more appropriate treatment for each individual patient. However, though high-throughput drug screening technology for this purpose is of the utmost importance in saving lives, there were many limitations to its wide use in many hospitals. The existing high-throughput drug combination screening technology consumes a large number of samples and consumes a considerable amount of expensive reagents. In addition, expensive automated liquid handlers, which were essential for exploring thousands of different pipetting, were not easy to introduce except for large-sized pharmaceutical companies and research institutes, which limited access to technology.
In this study, I construct a heterogeneous drug-loaded microparticle library by fabricating encoded photocurable polymer particle that has individually identifiable codes to track loaded drug. and I load various drug molecules, which I want to test to target cells, into each coded microparticle. Then, I developed to produce heterogeneous drug-laden microparticle arrays through simple self-assembly without the need for a microarray spotter or dispensing machine for generating microarray. I also have developed cell seeding method of seeding small-volume samples into the microwell-based cell chip. By utilizing the drug-laden microparticle hydrogel array and microwell-based cell chip technology, hundreds to thousands of different assays can be done at once with just a small number of samples and low cost.
Through the implemented platform, the anti-cancer drug sequential combination screening was conducted on the triple-negative breast cooler (TNBC) cells, which are generally known to be difficult to treat due to lack of known drug target, and the results of screening were analyzed by establishing a library of drugs in the EGFR inhibitory type and drugs in the genotoxin type. In addition, another study was conducted to find optimal drug combinations using patient-derived cells derived from tumors in patients with non-small cell lung cancer that have obtained acquired resistance. Finally, as the growing need for three-dimensional culture, such as spheroid and organoid for having a similar response to in vivo drug testing, it was also developed that microwell-based cell chip that is capable of 3D culture with low-cost and small-volume of cells.
The miniaturized in vitro anticancer drug screening platform presented in this study has the following significance. An easy-to-use technique that can be applied to a small number of patient cells or samples, which can dramatically reduce the use of conventional expensive equipment, reagents. The proposed technology in this study can be applied to a variety of academic studies previously inaccessible to high-throughput screening due to the high cost of reagents, the high price of equipment, or the limited amount of samples in conventional drug screening. and this platform can also dramatically increase access to clinical research in hospitals for personalized treatments. In particular, it is expected that the possibility of this platform will be further maximized if it is used in a relatively small and medium-sized research environment by the combined use of various rare samples such as patient-derived cells or patient-derived organoids.Chapter 1 Introduction οΌ
1.1 Motivation of this research οΌ
1.2 Competing technologies and Previous works οΌ
1.3 Main Concept: In vitro drug testing using miniaturized encoded drug-laden hydrogel array technology οΌοΌ
Chapter 2 Platform Development of Drug Releasing Hydrogel Microarray οΌοΌ
2.1 Encoded Drug-Laden Hydrogel & Library construction οΌοΌ
2.2 Array generation of heterogenous drug-laden microparticles. οΌοΌ
2.3 Cell Culturing on Cell Chip and bioassay οΌοΌ
Chapter 3 Sequential Drug Combination Screening Assy on TNBC οΌοΌ
3.1 Background : Sequential Drug Combination as promising therapeutic option οΌοΌ
3.2 Experimental design with sequential drug treatment assay οΌοΌ
3.3 Technical Issue & its engineering solution οΌοΌ
3.4 Assay Result οΌοΌ
Chapter 4 Drug Combination Assay on Patient-Derived Cells οΌοΌ
4.1 Background : Simultaneous Combination Treatment using Patient-Derived Cells οΌοΌ
4.2 Improvement of Platform for facilitating translational study οΌοΌ
4.3 Study Design for small-volume drug combinatorial screening with NSCLC patient derived cell οΌοΌ
4.4 Assay Result οΌοΌ
Chapter 5 Development of platform for 3D culture model οΌοΌ
5.1 3D culturable platform οΌοΌ
5.2 Development of 3D culture platform based Matrigel scaffold. οΌοΌ
5.3 Advantage over conventional 3D culture-based drug testing platform. οΌοΌ
Chapter 6 Conclusion οΌοΌ
Bibliography οΌοΌ
Abstract in Korean οΌοΌDocto