<논문 정보>
Ho Jin Han, Aneesh Sivaraman, Minkyoung Kim, Kyoung Ho Min, Mo Eun Song, Yongseok Choi, Won-Jun Choi, Hyo-Kyung Han, Junyeol Han, Jun-Pil Jang, In-Ja Ryoo, Kyeong Lee, Nak-Kyun Soung,
HIF-1α inhibition by MO-2097, a novel chiral-free benzofuran targeting hnRNPA2B1,
Journal of Advanced Research,
Volume 64,
2024,
Pages 67-81,
ISSN 2090-1232,
https://doi.org/10.1016/j.jare.2023.11.016.
(https://www.sciencedirect.com/science/article/pii/S2090123223003533)
Abstract: Introduction
Hypoxia-inducible factor 1 (HIF-1) is a transcriptional activator mediating adaptive responses to hypoxia. It is up-regulated in the tumor microenvironment and recognized as an effective anticancer drug target. Previously, we discovered that the natural compound moracin-O and its synthetic derivative MO-460 inhibited HIF-1α via hnRNPA2B1.
Objectives
This study aimed to develop novel HIF-1 inhibitors for cancer chemotherapy by harnessing the potential of the natural products moracins-O and P.
Methods
In an ongoing search for novel HIF-1 inhibitors, a series of nature-inspired benzofurans with modifications on the chiral rings of moracins-O and P were synthesized. They showed improved chemical tractability and were evaluated for their inhibitory activity on HIF-1α accumulation under hypoxic conditions in HeLa CCL2 cells. The most potent derivative's chemical-based toxicities, binding affinities, and in vivo anti-tumorigenic effects were evaluated. Further, we examined whether our compound, MO-2097, exhibited anticancer effects in three-dimensional cultured organoids.
Results
Herein, we identified a novel synthetic chiral-free compound, MO-2097, with reduced structural complexity and increased efficiency. MO-2097 exhibited inhibitory effects on hypoxia-induced HIF-1α accumulation in HeLa CCL2 cells via inhibition of hnRNPA2B1 protein, whose binding affinities were confirmed by isothermal titration calorimetry analysis. In addition, MO-2097 demonstrated in vivo efficacy and biocompatibility in a BALB/c mice xenograft model. The immunohistochemistry staining of MO-2097-treated tissues showed decreased expression of HIF-1α and increased levels of apoptosis marker cleaved caspase 3, confirming in vivo efficacy. Furthermore, we confirmed that MO-2097 works effectively in cancer patient-based organoid models.
Conclusion
MO-2097 represents a promising new generation of chemotherapeutic agents targeting HIF-1α inhibition via hnRNPA2B1, requiring further investigation.
Keywords: Nature-inspired chiral-free benzofuran; HIF-1α inhibition; hnRNPA2B1; RNA-binding protein; Patient-derived cancer organoid
- 배경
HIF-1
: 표적 유전자 전사를 자극하여 세포 증식, 혈관신생, 세포 에너지 대사, apoptosis 저항성, 전이와 같은 생물학적 과정 조절
저산소 상태에 빠진 경우 유도되는 단백질로 전사 인자로 기능한다.
저산소 환경이 혈관의 형성을 촉진하고 종양에서 혈관계의 형성에 중요하다.
| 구성 | 유전자 | 단백질 |
| HIF-1α | HIF1A | HIF-1, 알파 서브유닛 |
| HIF-1β | ARNT | ARNT |
| HIF-2α | EPAS1 | 내피 PAS 영역 단백질-1 |
| HIF-2β | ARNT2 | ARNT 2 |
| HIF-3α | HIF3A | HIF-3 알파 서브유닛 |
| HIF-3β | ARNT3 | ARNT 3 |
-method (biology)
cell culture: Hela CCL2 와 HCT116 세포주를 이용했다.
Apoptosis detection, Immunoblotting
semi-quantitative Rt-PCR
ITC, pull down assay 등의 실험 방법 이용
-결과
figure1.b
목적: 합성한 화합물의 세포성장에 미치는 영향
결과: 화합물 4-7은 세포 성장과 HIF-1 α 발현 감소
DMSO는 negative control, band가 4-7이 1-3 보다 희미한 것을 보아 발현이 감소됨을 알 수 있다.
marker가 베타엑틴-> cytosol에 존재한다.
figure. 2C
비오틴화 된 MO-2097을 이용하여 세포 추출물의 내인성 hnRNPA2B1의 선택적 pull down
hnRNPA2B1 외 다른 단백질과 cytosol marker인 Tubulin은 결합 X
따라서 두 단백질이 상호작용하고 있음을 알 수 있다.
figure. 6C
MO-2097이 hnRNPA2B1을 통해 HIF-1α를 억제함을 확인
MO-2097 처리 샘플에서 hnRNPA2B1과 HIF-1α 단백질의 변화를 관찰
오가노이드 4에서는 hnRNPA2B1의 발현이 높았고, HIF-1α 발현은 낮은 농도에서 반응함
- 결론
저산소 상태에서 새로운 벤조퓨란 화합물 MO-2097이 HIF-1α를 억제한다.
이러한 억제는 hnRNPA2B1에 의해 매개된다.
마우스 실험에서 독성 없이 효과 관찰됨
의의: RNA 결합 단백질이 항암제의 표적이 될 수 있다.
