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The Intracellular Delivery of Macromolecules by Modulating the Macropinocytosis Pathway /

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	The Intracellular Delivery of Macromolecules by Modulating the Macropinocytosis Pathway // Rabin Neupane.
作者:	Neupane, Rabin,
面頁冊數:	1 electronic resource (189 pages)
附註:	Source: Dissertations Abstracts International, Volume: 86-03, Section: B.
Contained By:	Dissertations Abstracts International86-03B.
標題:	Pharmacology. -
電子資源:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=31628959
ISBN:	9798384035510

The Intracellular Delivery of Macromolecules by Modulating the Macropinocytosis Pathway /
Neupane, Rabin,

The Intracellular Delivery of Macromolecules by Modulating the Macropinocytosis Pathway /Rabin Neupane. - 1 electronic resource (189 pages)

Source: Dissertations Abstracts International, Volume: 86-03, Section: B.

Drug molecules can reach their intracellular targets by different mechanisms, such as passive diffusion, transporter systems and endocytosis. Endocytosis is the process where cell internalize extracellular substrates by forming a vesicle and transporting it into the cell. In addition to the physiological role of endocytosis, it plays a vital role in the internalization of the therapeutics molecules. In the area of drug delivery, the most well investigated endocytic process are clathrin-mediated endocytosis, caveolar endocytosis and macropinocytosis. In addition to the conventional small therapeutic molecules, the advent of nano formulations and large molecules, such as nucleic acids, peptides and antibodies have broadened the landscape of drug delivery. Although the majority of the small therapeutic molecules undergo passive diffusion in cell, the large molecules and advanced targeted delivery systems, such as nanoparticles, are internalized into cell by endocytosis. Therefore, it is important to understand the endocytic routes in greater detail to improve the design of therapeutic molecules or formulations for successful delivery to intracellular targets. In this thesis, we aimed to investigate the feasibility of using novel macropinocytosis inducers to selectively induce macropinocytosis in cancer cells to increase the delivery of the therapeutic molecules.The test compounds were screened based on their capacity to increase the cellular uptake of dextran and albumin. Our results indicated that BAPT 78 (50 μM), BAPT 54 (50 μM) and MOMIPP (5 μM), significantly increased the uptake of dextran and albumin, in the colorectal cancer cells (CRC) lines, SW620 and HCT116. Dextran and albumin were internalized in cells by macropinocytosis, and the vacuoles were induced by the BAPT compounds (BAPT 78 and BAPT 54) and MOMIPP. Vacuoles formation was inhibited by the macropinocytosis inhibitors, cytochalasin D and ethyl isopropyl amiloride. These results were further validated by the morphological features of macropinocytosis (i.e., membrane ruffles formation and large endosomes > 0.5 μm), using transmission electron microscopy-based imaging. The vacuoles produced by BAPT 78 had late endosomal markers and interacted with lysosomes, indicating the endosomal compartment was acidic. The endolytic capacity of the lead compounds were low, suggesting that after the uptake of therapeutic molecules with aid of macropinocytosis, these molecules were susceptible to endolysosomal degradation. Furthermore, in the CRC cells, HCT116 and SW620, the uptake of an antibody, anti-β-tubulin, was increased by BAPT 78 (50 μM), BAPT 54 (50 μM) and MOMIPP (5 μM). The functional delivery of the plant toxin, saporin (10.9 to 350 nM) and small-interfering RNA (siRNA; 50 nM) to SW620 cells was very low after pre-incubating cells with the macropinocytosis inducers, BAPT 78 and MOMIPP. In contrast, the incubation of SW620, HCT116, SUM159 (mesenchymal triple-negative breast cancer) and a paclitaxel resistant variant of SUM159 triple-negative breast cancer cells (PAC200), with BAPT 78 (1.56 to 50 μM), BAPT 54 (1.56 to 50 μM) and MOMIPP (0.156 to 5 μM) significantly increased the uptake and functional delivery of the low molecular weight anticancer drugs, doxorubicin (6.2 to 200 nM) and paclitaxel (0.6 to 20 nM). Overall, our results suggest that by increasing endosomal escape, BAPT 78, BAPT 54 and MOMIPP, in combination with small molecules (doxorubicin and paclitaxel) and large molecules (saporin and siRNA), can increase uptake of these molecules, thereby increasing their intracellular concentrations in cancer cells to increase the efficacy of intracellular targeting.

English

ISBN: 9798384035510Subjects--Topical Terms:

634543
Pharmacology.
Subjects--Index Terms:

Macropinocytosis inducers

The Intracellular Delivery of Macromolecules by Modulating the Macropinocytosis Pathway /
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