Synthetic cytokine circuits that drive T cells into immune-excluded tumors

Allen, Greg M.; Frankel, Nicholas; Reddy, Nishith R.; Bhargava, Hersh K.; Yoshida, Maia A.; Stark, Sierra R.; Purl, Megan; Lee, Jungmin; Yee, Jacqueline L.; Yu, Wei; Li, Aileen W.; Garcia, K Christopher; El-Samad, Hana; Roybal, Kole T.; Spitzer, Matthew H.; Lim, Wendell A.

doi:10.1126/science.aba1624

Cited by 109 publications

(77 citation statements)

References 54 publications

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“…Our observations suggest that bystander killing mechanisms may enable CAR-T cells to target more tumor cells than would be predicted from the target antigen expression pattern, and thus may contribute to tumor destruction even in the presence of tumor heterogeneity. Similar or more potent bystander killing effects may be observed by arming CAR-T cells with cytokines such as IL-15(59,60) or IL-2(61), which may be safer than systematically administered cytokine therapy. It was estimated, for example, that under physiologic conditions in dense tissues such as lymph nodes, the cytokine IL-2, interacts over a characteristic length scale of 8-14 cell diameters (80-140 μm), which is determined by a balance between diffusion and consumption of cytokine(62).…”

Section: Discussionmentioning

confidence: 90%

Endogenous bystander killing mechanisms enhance the activity of novel FAP-specific CAR-T cells against glioblastoma

Ebert

Truong

et al. 2023

Preprint

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CAR-T cell therapies are being intensely investigated as a novel immunotherapy approach for glioblastoma (GBM), but so far clinical success has been limited. We recently described FAP as an ideal target antigen for GBM immunotherapy, with expression on tumor cells and tumor blood vessels occurring frequently in patients' tumors but with very limited expression in healthy tissues. Here, we generated a novel FAP-targeting CAR with CD3ζ and CD28 signaling domains and tested the resulting CAR-T cells for their ability to lyse GBM cells in vitro and in vivo. FAP-CAR-T cells showed target specificity against model cell lines and exhibited potent cytotoxicity against patient-derived glioma neural stem (GNS) cells. Remarkably, complete destruction of tumor cells was observed even where the antigen was expressed by a minor subpopulation of cells, indicating a bystander killing mechanism. Using co-culture assays, we confirmed the ability of FAP-CAR-T cells to mediate bystander killing of antigen-negative tumor cells, but only after activation by antigen-expressing target cells. This bystander killing effect was at least partially mediated by soluble factors. We also observed that the non-transduced fraction of the CAR-T cell product could be activated via T cell-secreted IL-2 to mediate antigen-non-specific killing, further amplifying the bystander effect. FAP-CAR-T cells controlled without overt toxicity the growth of subcutaneous tumors created using a mixture of antigen-negative and antigen-positive GBM cells. Together, our findings advance FAP as a leading candidate for clinical CAR-T cell therapy of GBM and highlight under-recognized antigen non-specific mechanisms that may contribute meaningfully to the antitumor activity of CAR-T cells.

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Section: Discussionmentioning

confidence: 90%

Endogenous bystander killing mechanisms enhance the activity of novel FAP-specific CAR-T cells against glioblastoma

Ebert

Truong

et al. 2023

Preprint

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“…We posit that adequate MR contrast would result from regional homing, expansion, and persistence of systemically delivered cells—a limitation of using NK-92 cells, which lack proliferation in animals due to irradiation prior to injection. Recent studies have emphasized an improvement in tumor infiltration, persistence, and expansion of infused cells when driving interleukin-2 ( 51 ) or a CAR upon SynNotch activation, which can both induce cell proliferation ( 32 ). We hope to incorporate similar strategies in the next iteration of our system with primary immune cells to boost activated cell detection sensitivity upon intravenous injection.…”

Section: Discussionmentioning

confidence: 99%

Visualizing cell–cell communication using synthetic notch activated MRI

Wang

Chen

Nyström

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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Cell–cell communication plays a fundamental role in multicellular organisms. Cell-based cancer immunotherapies rely on the ability of innate or engineered receptors on immune cells to engage specific antigens on cancer cells to induce tumor kill. To improve the development and translation of these therapies, imaging tools capable of noninvasively and spatiotemporally visualizing immune-cancer cell interactions would be highly valuable. Using the synthetic Notch (SynNotch) system, we engineered T cells that upon interaction with a chosen antigen (CD19) on neighboring cancer cells induce the expression of optical reporter genes and the human-derived, magnetic resonance imaging (MRI) reporter gene organic anion transporting polypeptide 1B3 (OATP1B3). Administration of engineered T cells induced the antigen-dependent expression of all our reporter genes in mice bearing CD19-positive tumors but not CD19-negative tumors. Notably, due to the high spatial resolution and tomographic nature of MRI, contrast-enhanced foci within CD19-positive tumors representing OATP1B3-expressing T cells were clearly visible and their distribution was readily mapped. We then extended this technology onto human natural killer-92 (NK-92) cells, observing similar CD19-dependent reporter activity in tumor-bearing mice. Furthermore, we show that when delivered intravenously, engineered NK-92 cells can be detected via bioluminescence imaging in a systemic cancer model. With continued work, this highly modular imaging strategy could aid in the monitoring of cell therapies in patients and, beyond this, augment our understanding of how different cell populations interact within the body during normal physiology or disease.

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“…A) Synthetic juxtacrine receptors (SJRs) are a class of synthetic receptors modular in both extracellular domain and intracellular domain. The extracellular domain consists of a choice ligand binding domain, and the intracellular domain consists of a transcription factor controlling a gene of choice 9,[15][16][17][18][19][20][21]35,36 . B) In the classical SJR circuit, the SJR direct activation circuit, the SJR binds its cognate ligand to release the transcription factor controlling the target gene.…”

Section: Design Of the Sjr Amplifier Circuitsmentioning

confidence: 99%

Design and Mathematical Analysis of Activating Amplifiers that Enable Modular Temporal Control in Synthetic Circuits

Lam

2023

Preprint

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The ability to control mammalian cells such that they self-organize or enact therapeutic effects as desired has incredible implications. Not only would it further our understanding of native processes such as development and the immune response, but it would also have powerful applications in medical fields such as regenerative medicine and immunotherapy. This control is typically obtained by synthetic circuits that use synthetic receptors, but control remains incomplete. For example, the synthetic juxtacrine receptors (SJRs) are widely used as they are fully modular and enable spatial control, but they have limited gene expression amplification and temporal control. I therefore designed transcription factor based amplifiers that amplify gene expression and enable unidirectional temporal control by prolonging duration of target gene expression. Using an in silico framework for SJR signaling, I combined these amplifiers with SJRs and show that these SJR amplifier circuits can improve the quality of self-organization and direct different spatiotemporal patterning. I then show that these circuits can improve chimeric antigen receptor (CAR) T cell tumor killing against two different types of tumors. These amplifiers are flexible tools that improve control over SJR based circuits and have both basic and therapeutic applications.

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Synthetic cytokine circuits that drive T cells into immune-excluded tumors

Cited by 109 publications

References 54 publications

Endogenous bystander killing mechanisms enhance the activity of novel FAP-specific CAR-T cells against glioblastoma

Endogenous bystander killing mechanisms enhance the activity of novel FAP-specific CAR-T cells against glioblastoma

Visualizing cell–cell communication using synthetic notch activated MRI

Design and Mathematical Analysis of Activating Amplifiers that Enable Modular Temporal Control in Synthetic Circuits

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