Hyaluronan-bile acid nanoparticles: Implications as a nanomedicine for the treatment of obesity-related chronic inflammatory diseases
- 주제(키워드) Self-assembled hyaluronic acid nanoparticles (HANPs) , Self-assembled Hyaluronan-Bile Acid nanoparticles (HABAs) , Chronic inflammatory diseases , Obesity , Psoriasis
- 주제(DDC) 547
- 발행기관 아주대학교
- 지도교수 김 욱
- 발행년도 2023
- 학위수여년월 2023. 2
- 학위명 박사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000032596
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Self-assembled hyaluronic acid nanoparticles (HANPs), consisting of the outermost hydrophilic HA layer surrounding multiple hydrophobic inner moieties, have been investigated broadly as a nanocarrier. Interestingly, several groups have recently reported that an empty HANP not bearing any drug is a potential therapeutic agent for the treatment of type 2 diabetes, arthritis, and atherosclerosis. Here, I report the efficacy of HANP in another chronic inflammatory diseases including obesity and psoriasis. First, I confirmed the potential of self-assembled hyaluronan-bile acid nanoparticles (HABAs) as therapeutics to treat obesity by assessing their in vitro and in vivo effects on adipogenesis and lipogenesis. Treatment of 3T3-L1 preadipocytes with HABA resulted in a dose-dependent suppression of adipogenesis and lipid accumulation, and decreased the expression of key adipogenic and lipogenic regulators. However, these HABA mediated effects were not observed in 3T3-L1 cells transfected with siRNAs against CD44. Consistent with in-vitro results, HABA treatment of diet-induced obese mice reduced the body weight and epididymal fat mass, and suppressed the induction of adipogenic and lipogenic regulators, while these effects were attenuated in the CD44-null mice. Subsequently, I investigated the potential of HABA as therapeutics for treating psoriasis. HABA exerted potent therapeutic efficacy against psoriasis-like skin by suppressing hyperproliferation of keratinocyte and M1 polarization of macrophage without overt toxicity signs. Transcutaneously administered HABAs were found to be accumulated and associated with pro-inflammatory M1 macrophages in the inflamed skin, and suppressed the M1 polarization of macrophage. As a result, HABA, compared with conventional drugs, revealed remarkable efficacy on IMQ-induced psoriasis-like mice. Taken together, I report that HANP have potential as a nanomedicine for treating obesity-related chronic inflammatory diseases.
more초록/요약
여러 소수성 중심 부분을 친수성 히알루론산으로 둘러싸고 있는 자가 조립 히알루론산 나노입자(HANP)는 나노 운반체로서 광범위하게 연구되어 왔다. 그런데 흥미롭게도 최근 몇 그룹에서 어떤 약물도 함유하지 않은 빈 HANP가 제2형 당뇨병, 관절염 및 동맥경화증 치료를 위한 잠재적인 치료제라고 보고했다. 여기, 나는 비만과 건선을 포함한 다른 만성 염증성 질환에서 HANP의 효능을 보고한다. 첫번째로 자가조립 히알루론-담즙산 나노 입자 (HABA)가 지방세포생성과 지방산합성에 미치는 효과를 평가하여 비만 치료제로서의 가능성을 확인했다. HABA를 3T3L-1 지방 전구세포에 처리하면 지방세포생성 및 지질 축적이 농도 의존적으로 억제되고, 주요 지방세포생성 및 지방산합성 조절인자의 발현이 감소했다. 그러나, 이러한 HABA 매개 효과는 CD44에 대한 siRNA로 형질주입 된 3T3-L1 세포에서는 관찰되지 않았다. 이러한 결과와 일치하게, 식이 유도 비만 마우스의 HABA 치료는 체중과 부고환 지방량을 감소시켰고, 지방세포생성 및 지방산합성 조절인자의 유도를 억제한 반면, CD44가 없는 마우스에서는 이러한 효과가 약화되었다. 그 다음으로 나는 건선 치료를 위한 치료제로서 HABA의 가능성을 확인했다. HABA는 뚜렷한 독성 징후 １５８ 없이 각질세포의 과증식과 대식세포의 M1 분화를 억제함으로써 건선 유사 피부에 대해 강력한 치료 효능을 발휘했다. 경피투여 된 HABA는 염증이 있는 피부에서 전 염증성 대식세포에 축적되어 대식세포의 M1 분극화를 억제하는 것으로 확인되었다. 그 결과, HABA는 IMQ로 유발된 건선 유사 마우스에서 기존 약물에 비해 현저한 효능을 보였다. 종합하면, 나는 HANP가 비만 관련 만성 염증성 질환 치료를 위한 나노의약으로서의 가능성을 가지고 있음을 보고한다.
more목차
PART I - Self-assembled hyaluronic acid nanoparticle suppresses fat accumulation via CD44 in diet-induced obese mice 1
I-1. INTRODUCTION 2
I-2. RESULTS 5
I-2.1 Synthesis and characterization of self-assembled HACN 5
I-2.2 Self-assembled HACNs suppress adipogenesis and lipogenesis during the 3T3-L1 differentiation 10
I-2.3 HACN mediated effects on adipogenesis and lipogenesis depend on CD44 15
I-2.4 In vivo effects of HACN in WT and CD44 KO DIO mice 20
I-2.5 Effects of CA and free HA on adipogenesis and weight change 26
I-3. DISCUSSION 31
PART II - Hyaluronic Acid Nanoparticles as a Topical Agent for Treating Psoriasis 33
II-1. INTRODUCTION 34
II-2. RESULTS 37
II-2.1. Synthesis and characterization of self-assembled HALN 37
II-2.2. Therapeutic efficacy of HALN administered via the subcutaneous route in psoriasis-like skin dermatitis 47
II-2.3. Accumulation of Transcutaneously Administered HALN in IMQ-Inflamed Dermis 63
II-2.4. Efficacy of Transcutaneously Administered HALNs against Psoriasis-like Skin Dermatitis 80
II-2.5. Macrophages as a Target in IMQ-Inflamed Skin 97
II-2.6. Modulation of Macrophage Polarization 104
II-3. DISCUSSION 117
III. MATERIALS and METHODS 120
2.1. Materials 120
2.2. Characterization of Compounds 120
2.3. CA Ester 120
2.4. EtCA 121
2.5. HA-CA 122
2.6. LCA Ester 122
2.7. EtLCA 123
2.8. HA-LCA 124
2.9. 10k-HA-LCA 124
2.10. Cy5.5-HA 125
2.11 Cy5.5-HA-LCA and Cy5.5-HA-CA 125
2.12. Characterization of HANPs 126
2.13. The 3T3-L1 cell culture and differentiation 126
2.14. The 3T3-L1 cell viability assay 127
2.15. Transfection of small tnerfering RNA 128
2.16. Mice 128
2.17. Histology and Immunofluorescence 128
2.18. Quantitative real-time PCR analysis 130
2.19. High-fat diet induced obesity mice 132
2. 20. Determination of adipocyte size 132
2.21. Oil-red-O staining 133
2. 22. Colorimetric Assessment of Terminal N-Acetyl-d-Glucosamine (Hyaluronan Breakdown Products) 133
2. 23. Psoriasis-like Mouse Models 134
2.24. Cell Isolation from the Shaved Dorsal Skin 136
2.25. Hematological Toxicity 136
2.26. In Vivo Toxicity Test 137
2.27. In Vivo Skin Penetration Studies 137
2.28. Multi-Photon Microscopy 128
2.29. Ex Vivo Biodistribution 128
2.30. Cell Culture 139
2.31. In Vivo Depletion of Macrophages 140
2.32. In Vitro Competitive Inhibition Assay 140
2.33. In Vitro Cellular Uptake Assay 141
2.34. RNA-seq and Bioinformatic Analysis 141
2.35. Immunoblot Analysis 142
2.36. In Vitro Macrophage Polarization 143
2.37. Statistical analysis 144
IV. CONCLUSION 146
V. REFERENCE 147
VI. ABSTRACT IN KOREAN (국문초록) 157
