Generation and testing of clinical-grade exosomes for pancreatic cancer
Mayela Mendt, … , Valerie S. LeBleu, Raghu Kalluri
Mayela Mendt, … , Valerie S. LeBleu, Raghu Kalluri
Published April 19, 2018
Citation Information: JCI Insight. 2018;3(8):e99263. https://doi.org/10.1172/jci.insight.99263.
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Research Article Oncology

Generation and testing of clinical-grade exosomes for pancreatic cancer

Abstract

Exosomes are extracellular vesicles produced by all cells with a remarkable ability to efficiently transfer genetic material, including exogenously loaded siRNA, to cancer cells. Here, we report on a bioreactor-based, large-scale production of clinical-grade exosomes employing good manufacturing practice (GMP) standards. A standard operating procedure was established to generate engineered exosomes with the ability to target oncogenic Kras (iExosomes). The clinical-grade GMP iExosomes were tested in multiple in vitro and in vivo studies to confirm suppression of oncogenic Kras and an increase in the survival of several mouse models with pancreatic cancer. We perform studies to determine the shelf life, biodistribution, toxicology profile, and efficacy in combination with chemotherapy to inform future clinical testing of GMP iExosomes. Collectively, this report illustrates the process and feasibility of generating clinical-grade exosomes for various therapies of human diseases.

Authors

Mayela Mendt, Sushrut Kamerkar, Hikaru Sugimoto, Kathleen M. McAndrews, Chia-Chin Wu, Mihai Gagea, Sujuan Yang, Elena V. Rodriges Blanko, Qian Peng, Xiaoyan Ma, Joseph R. Marszalek, Anirban Maitra, Cassian Yee, Katayoun Rezvani, Elizabeth Shpall, Valerie S. LeBleu, Raghu Kalluri

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Figure 1

Specific targeting of iExosomes for human PDAC cells with KrasG12D.

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Specific targeting of iExosomes for human PDAC cells with KrasG12D. (A) ...
(A) Volcano plots depicting log2 fold change (red, upregulated genes; blue, downregulated genes; gray, genes that were not significantly deregulated) and –log10 (P value) of differentially expressed genes between siKrasG12D–1 iExo–treated Panc-1 cells and all controls and BxPC-3 cells. (B) Tumor volume measured by MRI of siKrasG12D–1 iExo (n = 7) or siScrbl iExo (n = 7) at baseline (day 0 = day 62 after tumor induction) and after treatment (day 30, 81, and 228 posttreatment start [PTS]). #, no measurement available; mice died. (C) Representative MRI images of PDX pancreas tumors; a yellow dashed line encircles the tumors. (D) Kaplan-Meier curve indicating the survival of PDX mice in the listed treatment groups after birth (siKrasG12D–1 iExo [n = 7], siScrbl iExo [n = 7]; log-rank (Mantel-Cox) test). The approximated time for the natural life span of nude mice is indicated by the red dotted line. (E) Tumor weight at end point in studies with PDX mice (siKrasG12D iExo [n = 7], siScrbl iExo [n = 7]). (F) Representative H&E image of aggressively invasive pancreatic tumors in siScrbl iExo–treated mice, in comparison with predominant inflammation and markedly reduced tumors in pancreata of siKrasG12D–1 iExo–treated mice. Scale bars: 100 μm (left); 50 μm (right). The data are presented as the mean ± SEM. Unless otherwise stated, unpaired 2-tailed t test was used to determine statistical significance. *P < 0.05, ** P < 0.01. See Supplemental Source Data 1 and 2.