Bioinstructive implantable scaffolds for rapid in vivo manufacture and release of CAR-T cells

Agarwalla, Pritha; Ogunnaike, Edikan A.; Ahn, Sarah; Froehlich, Kristen; Jansson, Anton; Ligler, Frances S.; Dotti, Gianpietro; Brudno, Yevgeny

doi:10.1038/s41587-022-01245-x

Cited by 94 publications

(86 citation statements)

References 53 publications

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“…Nevertheless, our discovery suggests that their formation is also regulated by the endothelium and specifically by Notch activity in hemogenic cells. This new information could be used to optimize engineering T cells during ex-vivo production, for example during CAR-T cells manufacture ( 59 ).…”

Section: Discussionmentioning

confidence: 99%

Hematopoietic stem and progenitor cell heterogeneity is inherited from the embryonic hemogenic endothelium

Ghersi

Baldissera

Hintzen

et al. 2022

Preprint

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Multipotent hematopoietic stem/progenitor cells (HSPCs) generate all mature blood cells in the erythroid, lymphoid, and myeloid lineages. HSPCs are initially produced in the embryo, via transdifferentiation of hemogenic endothelial cells (hemECs) in the aorta-gonad mesonephros (AGM). HSPCs in the AGM are functionally heterogenous in differentiation and proliferative output, but how these intrinsic differences are acquired remains unanswered. This knowledge could inform approaches to overcome the dysregulation of HSPC heterogeneity associated with poor outcomes of autologous transplants. Here we discovered that loss of microRNA (miR)-128 (miR-128Δ/Δ) in zebrafish leads to an expansion of hemECs forming replicative HSPCs in the AGM, and a skew towards the erythroid and lymphoid lineages in larval and adult stages. Furthermore, we found that inhibiting miR-128 during the differentiation of human pluripotent stem cells into hemECs, but not during the endothelial-to-hematopoietic transition, recapitulated the lineage skewing. In vivo, expression of wild-type miR-128 in endothelium restored the blood lineage distribution in miR-128Δ/Δ zebrafish. We found that miR-128 represses the expression of the Wnt inhibitor csnk1a1 and the Notch ligand jag1b, and thus promotes Wnt and Notch signaling in hemECs. De-repression of cskn1a1 resulted in hemECs generating replicative and erythroid-biased HSPCs, whereas de-repression of jag1b resulted in hemECs forming lymphoid-biased HSPCs in the AGM and relative mature blood cells in adult. We propose that HSPC heterogeneity is established in hemogenic endothelium prior to transdifferentiation and is programmed in part by Wnt and Notch signaling modulation.

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Section: Discussionmentioning

confidence: 99%

Hematopoietic stem and progenitor cell heterogeneity is inherited from the embryonic hemogenic endothelium

Ghersi

Baldissera

Hintzen

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…From a proof-of-concept study by Agarwalla, P and colleagues using modified alginate scaffolds loaded with retroviral vectors containing CD19 CAR-encoding genes, in vitro results demonstrated an equivalent amount of CAR transduced cells compared to the conventional approach as well as a non-inferior capability in tumor cell eradication. In vivo generation of CAR T cells using subcutaneous implants in tumor xenograft mice showed sufficient tumor-lytic effects on CD19 + tumor cells, and mice exhibited an equal survival rate to those treated with conventional ACT therapy [119].…”

Section: Challenges and Future Perspectivesmentioning

confidence: 96%

Potential roles of hyaluronic acid inin vivoCAR T cell reprogramming for cancer immunotherapy

et al. 2022

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“…Similarly, pre-clinical efforts have resulted in the in vivo generation of fibroblast activation protein (FAP)-directed CAR-T cells to treat cardiac injury, by selectively reprogramming T cells using a CD5-targeted LNP system ( 193 ). Besides LNP-mediated strategies, implantable bioinstructive scaffolds have been developed, which can generate CAR-T cells in vivo and, hence, circumvent the cost-intensive and lengthy vein-to-vein times required for ex vivo manufacturing of autologous CAR-T products ( 194 ).…”

Section: In Vivo Gene Editing and Cell Engineering Strategiesmentioning

confidence: 99%

Multiplexed engineering and precision gene editing in cellular immunotherapy

2022

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The advent of cellular immunotherapy in the clinic has entirely redrawn the treatment landscape for a growing number of human cancers. Genetically reprogrammed immune cells, including chimeric antigen receptor (CAR)-modified immune effector cells as well as T cell receptor (TCR) therapy, have demonstrated remarkable responses across different hard-to-treat patient populations. While these novel treatment options have had tremendous success in providing long-term remissions for a considerable fraction of treated patients, a number of challenges remain. Limited in vivo persistence and functional exhaustion of infused immune cells as well as tumor immune escape and on-target off-tumor toxicities are just some examples of the challenges which restrain the potency of today’s genetically engineered cell products. Multiple engineering strategies are being explored to tackle these challenges.The advent of multiplexed precision genome editing has in recent years provided a flexible and highly modular toolkit to specifically address some of these challenges by targeted genetic interventions. This class of next-generation cellular therapeutics aims to endow engineered immune cells with enhanced functionality and shield them from immunosuppressive cues arising from intrinsic immune checkpoints as well as the hostile tumor microenvironment (TME). Previous efforts to introduce additional genetic modifications into immune cells have in large parts focused on nuclease-based tools like the CRISPR/Cas9 system or TALEN. However, nuclease-inactive platforms including base and prime editors have recently emerged and promise a potentially safer route to rewriting genetic sequences and introducing large segments of transgenic DNA without inducing double-strand breaks (DSBs). In this review, we discuss how these two exciting and emerging fields—cellular immunotherapy and precision genome editing—have co-evolved to enable a dramatic expansion in the possibilities to engineer personalized anti-cancer treatments. We will lay out how various engineering strategies in addition to nuclease-dependent and nuclease-inactive precision genome editing toolkits are increasingly being applied to overcome today’s limitations to build more potent cellular therapeutics. We will reflect on how novel information-rich unbiased discovery approaches are continuously deepening our understanding of fundamental mechanisms governing tumor biology. We will conclude with a perspective of how multiplexed-engineered and gene edited cell products may upend today’s treatment paradigms as they evolve into the next generation of more potent cellular immunotherapies.

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Bioinstructive implantable scaffolds for rapid in vivo manufacture and release of CAR-T cells

Cited by 94 publications

References 53 publications

Hematopoietic stem and progenitor cell heterogeneity is inherited from the embryonic hemogenic endothelium

Hematopoietic stem and progenitor cell heterogeneity is inherited from the embryonic hemogenic endothelium

Potential roles of hyaluronic acid inin vivoCAR T cell reprogramming for cancer immunotherapy

Multiplexed engineering and precision gene editing in cellular immunotherapy

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