Reprogrammed: A biotechnology Podcast

This episode looks at two papers, one a 2019 paper by Hassett et al. about optimizing Lipid Nanoparticles specifically for intramuscular mRNA vaccines. And a second, Laczko et al., from 2020, that shows what kind of immune responses you can get from nucleoside-modified mRNA-LNP after a single shot in mice. Together, these explain why mRNA vaccines aren’t just ‘RNA magic.’ They’re a delivery story, and an immune-programming story.

This episode discusses the 2005 Kariko et al. paper about RNA immunogenicity.

In this episode, we explain why influenza is such a persistent threat—its segmented RNA genome enables antigenic drift and reassortment-driven “shift,” letting it change faster than traditional vaccines can be manufactured. We thenwalk through Petsch et al. (2012), showing that synthetic mRNA vaccines elicit functional anti-HA antibodies measured by HAI, protect mice from lethal influenza challenge (with protection transferable by antibody), and cangenerate remarkably durable immunity—highlighting the real breakthrough: turning vaccine development from slow biological manufacturing into speed-of-information medicine.

mRNA vaccines: when the call is coming from inside the house, discusses how the immune system knows what's going on inside of cells and how specific that knowledge is. This episode explains MHC Haplotype and how MHC restriction limits the targets of CTLs.

This episode explores an early mRNA vaccination study showing that injected RNA can be injected into the body, be translated into protein that is presented on MHC class I to generate an antigen-specific cytotoxic T-cell respons. This discussion highlights how researchers moved from simply proving in-vivo protein expression to demonstrating that nucleic acids themselves could function as immunogens—laying the conceptual groundwork for modern mRNA vaccines.

This week on Reprogrammed, Jack and Annie tackle a key limitation in CAR T cell therapy for solid tumors: poor trafficking to the tumor site. Centered on the 2010 study by Craddock et al., this episode explores how engineering T cells to express the chemokine receptor CCR2b—matching the tumor’s secretion of CCL2—can dramatically improve tumor infiltration without sacrificing cytotoxic function. Through in vitro migration assays and in vivo imaging in mouse models, the team unpacks how this strategy adds a modular “navigation” system to CAR T cells, allowing them to home in on their targets more effectively.With references to past episodes on co-stimulatory signaling, persistence, and dual-antigen targeting, this episode expands the design philosophy of CAR T therapy from “stronger” to “smarter.” Annie and Jack also look ahead to next week’s discussion of split-signal CARs that promise greater safety through precision control—bringing us closer to truly programmable immunity.

In this episode, we examine the work of Roselli et al.1, who explore a critical frontier in CAR T cell engineering: how to build T cells that don’t just kill tumors effectively, but also survive, persist, and adapt in the complex and hostile environments characteristic of solid tumors and relapsing hematologic malignancies.Building on themes from earlier episodes—such as the impact of co-stimulatory domains (CD28 vs. 4-1BB), strategies to reduce tonic signaling (Eyquem et al.), and optimizing cytokine support (IL-15)—this paper asks: Can we combine the best aspects of multiple co-stimulatory signals into a single CAR design? And, can dual-antigen targeting reduce the likelihood of tumor escape?Roselli et al. engineer several new CARs that incorporate both 4-1BB (for metabolic fitness and longevity) and a modified version of CD28 (mut06) designed to reduce exhaustion while retaining strong activation. They also explore tandem CARs targeting both CD19 and CD20, to limit the risk of antigen escape—a known challenge with CD19-directed therapies.Using the xCelligence platform, in vivo models, and transcriptional profiling, they show that these dual co-stimulatory CARs outperform their single-signal counterparts in terms of cytotoxicity, expansion, cytokine production, and resistance to exhaustion. The final experiments confirm that dual-antigen targeting not only delays relapse but also increases T cell persistence and flexibility, especially in tumors with heterogeneous antigen expression.In the broader arc of this podcast, this paper illustrates the field’s move toward rational CAR design and continuous improvement—combining lessons from receptor architecture, signaling dynamics, immune memory, and tumor evolution to create therapies that are both powerful and durable.

Over the past episodes of Reprogrammed, we’ve followed the rapid evolution of CAR T cell therapy—from early experiments demonstrating that antibody-based chimeric receptors could trigger T cell activation and IL-2 production, to the incorporation of co-stimulatory domains like CD28 and 4-1BB that dramatically improved T cell survival and cytotoxic efficacy.In this episode of Reprogrammed, Jack and Annieexplore a pivotal 2014 Science paper by Tran et al.,which presents a landmark case in personalized cancer immunotherapy. Shifting focus from engineered CAR T cells to the body's natural immune capabilities, the study highlights how mutation-specific CD4⁺ T cells derived from a patient with metastatic cholangiocarcinoma were isolated, expanded, and infused back to achieve significant tumor regression. The episode delves into the biology of helper T cells, the use of tandem minigene screening to identify a neoantigen in the ERBB2IP gene, and how these T cells exhibited a potent Th1 response, directly killing tumor cells. Jack and Annie discuss the therapeutic promise and the technical and logistical limitations of such personalized approaches, framing the work as both a scientific milestone and a reminder of the untapped potential within the immune system itself.

Over the past episodes of Reprogrammed, we’ve followed the rapid evolution of CAR T cell therapy—from early experiments demonstrating that antibody-based chimeric receptors could trigger T cell activation and IL-2 production, to the incorporation of co-stimulatory domains like CD28 and 4-1BB that dramatically improved T cell survival and cytotoxic efficacy.In episode 5, we saw how CD28 co-stimulation, when engineered into CARs, could prevent Activation-Induced Cell Death (AICD) and promote proliferation. Then, in episode 6, we contrasted this with 4-1BB-based signaling, which provided a more durable survival signal and fostered memory-like cell states. Most recently, we saw how cytokines like IL-15 could be used to extend the persistence and antitumor activity of engineered T cells in vivo.What all these strategies shared was a focus on enhancing T cell function and persistence by manipulating the CAR’s intracellular signaling domains or the cell’s surrounding environment.But Eyquem et al. (2017) raised a new question entirely, i.e., does transfecting these T Cells pose an unaccounted for risk of causing unwanted effects as a result of where in the T Cell genome the CAR construct was integrated?

Reprogrammed is a biotechnology podcast exploring the development of cell therapies, with this season focusing on CAR T cells in cancer treatment. The Brentjens et al. (2013) study marked a pivotal moment in the clinical application of CAR T therapy, demonstrating for the first time that second-generation CD19 CAR T cells incorporating a CD28 co-stimulatory domain could induce molecular remission in adults with relapsed/refractory B-ALL. By bridging the gap between preclinical engineering and real-world application, this study laid the foundation for the eventual FDA approval of CD19-directed CAR T therapies for B-cell malignancies

This podcast delves into the exciting world of cancer immunotherapy, focusing on the evolution of cancer vaccines and the recent surge of cell-based therapies. This series follows a string of primary research papers highlighting the most impactful advances in CAR T Cell therapy. It is a companion to a KU Edwards Campus Biotechnology course, but it contains information interesting to anyone who wants to understand the mechanics of this therapy. Like the last episode, this paper endeavors to solve the problem of Activation-induced cell death (AICD) amongst recombinant, antigen-directed T Cells. Both papers considered CD28 co-stimulation by B7.1, with Maher et al electing for CD28 incorporation into the antigen receptor, and this paper relying on B7 expression by target cells. This paper also incorporates the addition of IL-15 as supportive of the long-term survival of the T Cells.

This podcast delves into the exciting world of cancer immunotherapy, focusing on the evolution of cancer vaccines and the recent surge of cell-based therapies. This series follows a string of primary research papers highlighting the most impactful advances in CAR T Cell therapy. It is a companion to a KU Edwards Campus Biotechnology course, but it contains information interesting to anyone who wants to understand the mechanics of this therapy. It begins with a 2003 Nature Biotechnology paper by Maher et al. that combines CD3 and CD28 signaling domains together on a single chimeric antigen receptor. A second paper from the 2004 journal, Leukemia, by Imai et al., looks at another receptor’s signaling domain, 4-1BB, as an alternative strategy to overcoming AICD in CAR T Cells.

This podcast delves into the exciting world of cancer immunotherapy, focusing on the evolution of cancer vaccines and the recent surge of cell-based therapies. This series follows a string of primary research papers highlighting the most impactful advances in CAR T Cell therapy. It is a companion to a KU Edwards Campus Biotechnology course, but it contains information interesting to anyone who wants to understand the mechanics of this therapy. It begins with a 1998 Journal of Experimental Medicine article by Krause et al. that details the creation of a chimeric receptor, 3G6-CD28, that incorporates the signaling domain of CD28 in the cytoplasmic portion of the antigen receptor to enhance the survival and proliferation of T lymphocytes targeting tumor cells. Experiments using Jurkat cells and human primary T cells demonstrate that the 3G6-CD28 chimeric receptor triggers CD28 signaling upon GD2 recognition, improving T cell survival and expansion, especially in the presence of anti-CD3 stimulation. This approach offers a potential strategy for improving cell-based immunotherapy by overcoming the limitations of tumor cells' poor antigen presentation and lack of costimulatory molecules. The results show a significant improvement in T-cell activity against tumor cells.

This is the first paper to demonstrate that plasmid DNA injections elicited humoral immune responses and could therefore function as vaccines.

This episode addresses the question of whether lymphocytes can be harvested, reprogrammed in their specificity for a selected tumor antigen ex vivo, and then re-administered to the patient to combat their cancer.  The first paper, from 1989, by Gross et al., examines whether a chimeric receptor, forged from the antigen-binding region of an immunoglobulin and the internal signaling elements of a T Cell Receptor, could be functionally expressed on harvested cells. A review from 2023, will provide context for how this nascent work has since developed into a powerful, clinical therapy against some forms of cancer.

This episode addresses the question of whether lymphocytes directed against one specific tumor antigen can be manipulated to better combat cancer. The first paper, from 1989, examines cells initially generated in an animal, but then be isolated, activated ex vivo and readministering to the animal to see if they will effectively fight against a tumor that was previously not recognized by the immune system. The second paper, from 2002, looks specifically at the capacity to use CD8+‘killer’ T Cells in the clinic as an anti-melanoma therapy.

Introduction of key papers:o 1974 article by Israel Penn, MD, on cancer in immune deficiencies.2020 article by Hegde et al. on dendritic cells and immune surveillance.• Framing question: “Does the immune system operate to protect us against cancer?”

This podcast delves into the exciting world of cancer immunotherapy, focusing on the evolution of cancer vaccines and the recent surge of cell-based therapies. It begins with a brief historical overview, highlighting the discovery of Coley’s toxin, which marked the earliest attempts at cancer vaccination. It then dives into the complexities of therapeutic vaccines, underscoring the challenges presented by cancer cells' ability to evade the immune system. The conversation transitions to cell-based therapies, which involve extracting and modifying immune cells outside the body to target cancer cells directly. It then highlights CAR T Cell therapy’s effectiveness against blood cancers and the ongoing research to improve its efficacy against solid tumors. Then it delves into the potential of IL-15, a cytokine that seems to enhance CAR T-cell function and longevity, and explores the role of NK cells as a potential alternative platform for CAR engineering. Finally, the dialogue addresses the crucial ethical considerations surrounding the rapid advancement of personalized medicine and the importance of ensuring equitable access to these transformative therapies. The purpose is to educate the audience about the evolution of cancer immunotherapy and the potential of cell-based therapies to revolutionize cancer treatment. 

This episode relates to: the 1990 Wolff et al., paper in Science,247(4949 Pt 1):1465–1468. Direct Gene Transfer into Mouse Muscle in Vivoandthe 1993 Ulmer et al., paper in Science, 259(5102):1745–1749. Heterologous protection against influenza by injection of DNAencoding a viral protein.

This podcast refers to a paper by Wang et al., PNAS,90(9):4156–4160. Gene inoculation generates immune responses against human immunodeficiency virus type 1. 1993.