Longitudinal Monitoring of Ribonucleic Acid Content of a Live Cell with a Nanotube Pore Interface

Project Overview

This project aimed to develop an approach for interfacing carbon nanotube porins (CNTPs) to live cell membranes and using this interface to extract and identify genetic information. The initial experiments led to successful establishment of such interface; however, during these experiments our team discovered an unexpected effect where the CNTP-cell interface proceeded further to show signs of membrane fusion. Subsequent experiments confirmed this initial assessment and indicated that CNTP-cell interfaces would indeed preferentially proceed toward fusion. These observations opened up an opportunity to develop this phenomenon into a direct cytoplasmic drug delivery approach. During the second phase of the project, we have demonstrated that CNTP-studded liposomes can effectively fuse to other liposomal subpopulations. This process is driven a novel fusion mechanism catalyzed by a CNTP dimer spontaneously formed in the membrane. The project team also demonstrated that this fusion mechanism is effective in live cells. Our experiments showed that when our CNTP-studded liposomes carry a cargo of a chemotherapy drug, they can deliver this drug to several types of cancer cells, killing them efficiently. We demonstrated that the efficiency of our approach was comparable to conventional chemotherapy delivery approaches but used a significantly lower (and thus less systemically toxic) drug dose. Thus, the main technical outcome of this project was the development of a novel drug delivery approach and its demonstration in cancer cell cultures. The results of this research have been published in high-impact journal (1) and have been filed as a provisional US/PCT patent application (2).

Mission Impact

This work has developed, patented, and published a new technological approach for delivery of liposome-encapsulated cargo to live cells. The project demonstrated efficient delivery of a chemotherapy agent, doxorubicin, to several types of cancer cells. These experiments indicate that this delivery technology is promising for direct cytoplasmic delivery of anticancer drugs, vaccines, and genetic materials. The PI has already submitted two follow-on grant proposals to external sponsors to develop these capabilities further. Overall, this technology contributes to LLNL's core competency in Bioscience and Bioengineering, and specifically to development of more effective countermeasures and methods for their delivery.

Publications, Presentations, and Patents

N.T. Ho, M. Siggel, K.V. Camacho, R.M. Bhaskara, J.M. Hicks, Y-C. Yao, Y. Zhang, J. Köfinger, G. Hummer, A. Noy. "Drug delivery with carbon nanotube porin fusogens," Proc. Natl. Acad. Sci. USA, v. 118 (19), p. e2016974118 (2021).

A. Noy, N. Ho, G. Hummer, M. Siggel. "Nanotube-vesicle compositions and uses thereof." US Provisional 62/948,169, 2019.