[CCoE Notice] Cullen College Dissertation Defense Announcement - Arash Saeedi

[ccoecomm at Central.UH.EDU]

Rachel Knudsen
Executive Director of Communications
Cullen College of Engineering
Engineering Building 2, Suite E311
University of Houston
Houston, TX 77204-4007
713.743.4379 – riward at central.uh.edu<mailto:riward at central.uh.edu>
________________________________
From: Thomas, Yolanda M <YMThomas at Central.UH.EDU>
Sent: Tuesday, November 29, 2022 9:53 AM
To: Knudsen, Rachel W <riward at Central.UH.EDU>
Subject: Cullen College Dissertation Defense Announcement - Arash Saeedi


[Dissertation Defense Announcement at the Cullen College of Engineering]

Engineering Non-cytotoxic Delivery of Proteins by T cells via Fusion to NPC2



Arash Saeedi

December 5, 2022;  1:00 PM – 3:00 PM (CST)

Zoom Link: https://urldefense.com/v3/__https://uh-edu-cougarnet.zoom.us/j/94533319289?pwd=WG5SWERERjhuR3hLSTRUOUxCYjZyZz09__;!!LkSTlj0I!Buij65ujGg04jios5e70p0sGRMkvH_Hbz8exO1YFN2ty7cNKUBPHy0MaDOLr1T8aYvOleYR0bx_DV9JgOnEKulwoDXs$ <https://urldefense.com/v3/__https:/uh-edu-cougarnet.zoom.us/j/94533319289?pwd=WG5SWERERjhuR3hLSTRUOUxCYjZyZz09__;!!LkSTlj0I!FscYlfSmUu9eq4kmTgrHaAQngpD3XqtQ-vBOqUqFxQy7-ZmOf5Ux0voJjqxRViDmV7qADn1Ee94oeou12HFTi-L2XA$>

Meeting ID: 945 3331 9289
Passcode: 732852



Committee Chair:
Navin Varadarajan, PhD

Committee Members:
Richard Willson, Ph.D. | Mehmet Orman, Ph.D. | Weiyi Peng, Ph.D. | Fatima Merchant, PhD.

Abstract

      Engineering cellular therapeutics by programming T cells has great potential in immunology. The primary mechanism employed by T cells for the specific transfer of proteins at the immunological synapse is via the lysosomal perforin pathway that facilitates the transfer of cytotoxic granzymes leading to apoptosis in target cells. Facilitating the delivery of non-cytotoxic proteins through perforin oligomers will dramatically expand the range of protein cargos that T cells can traffic to the target cells. Here, we have identified the intralysosomal protein, NPC2, as a chaperone that can facilitate the delivery of T-cell derived reporter proteins through perforin pores at the immunological synapse. Structural and biophysical considerations suggested that NPC2 could traverse through perforin pores and in vitro experiments confirmed the transport of purified NPC2 through perforin pores on cell membranes. To characterize the ability of NPC2 to facilitate the transfer of payloads in T cells, we constructed NPC2-mCherry fusion proteins in T cells. Using confocal microscopy and flow cytometry, we confirmed the colocalization of the NPC2 fused protein with lytic granules and the transfer of the fluorescent protein payload from T cells to target cells in co-culture experiments. The NPC2 fusion enabled the localization of mCherry to secretory lysosomes in mouse TCR CD8+ T cells and human CD4+ and CD8+ chimeric antigen receptor (CAR) T cells.  Finally, we introduce a novel method for expression of NPC2 fused toxins within CAR T cells for treating immune resistant tumor cells. These results illustrate that by using NPC2 as a molecular chaperone, the NPC2-perforin pathway can be exploited as a programmable molecular delivery system for cell-based therapies.

[Engineered For What's Next]


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