Translational Science Techniques

Researchers at Johns Hopkins University have created a revolutionary protein “switch” that tricks cancer cells into manufacturing their own chemotherapy drugs, causing them to self-destruct while sparing healthy cells. Instead of delivering drugs directly to cancer cells, this method uses a harmless “prodrug” that only becomes activated inside cancer cells when the switch detects specific cancer markers. The switch is made by combining two proteins: one that senses cancer markers and another from yeast that converts the inactive prodrug into a potent cancer-killing drug. When the switch detects cancer, it activates the drug inside that cell, turning the cancer cell into a drug factory that destroys itself. To work, the switch must enter cancer cells either by delivering the protein itself or by inserting the gene that makes the protein, allowing the cancer cell’s own machinery to produce the switch. Afterward, patients receive the inactive chemotherapy prodrug, which becomes activated only inside cancer cells. This new approach focuses on producing the drug inside cancer cells rather than just delivering it to them, which could kill more cancer cells while reducing harmful side effects on healthy tissue. Lab tests on human colon and breast cancer cells have shown promise, and animal testing is expected to start within a year. While still early, this technique offers a radically different way to attack cancer.

Remember The Jetsons - that futuristic cartoon where technology could deliver anything you needed? That’s exactly what comes to mind when I think about antibody drug conjugates (ADCs) They’re like Uber Eats for cancer therapy They have a programmed delivery address (the antibody component that targets a specific tumor antigen) and a payload to drop off at that address (the cytotoxic agent) No wandering around. No collateral damage (in theory). Just targeted delivery. Meet the newest ADC on the block: datopotamab deruxtecan The delivery address for this ADC are cells that express TROP2 (trophoblast cell surface antigen 2) TROP2 activates pathways for self-renewal, proliferation, invasion, and survival, and has been associated with drug resistance. It’s expressed in many normal cells but overexpressed in malignant cells. (this is the same address sacituzumab govitecan (Trodelvy) delivers to) What’s in the trunk of this ADC? The payload is deruxtecan — a topoisomerase I inhibitor (the same payload in trastuzumab deruxtecan (Enhertu)) Topoisomerases are enzymes that help unwind DNA so it can replicate Blocking that process is like blowing up the kitchen where the cell is cooking up new DNA 💣 No kitchen = no replication = cell death Datopotamab deruxtecan was recently approved for metastatic HR+/HER2- breast cancer based on the TROPION-Breast01 study Patients were randomized to datopotamab deruxtecan (n=365) or investigator’s choice of chemotherapy (n=367), which included: eribulin (60%), capecitabine (21%), vinorelbine (10%), or gemcitabine (9%). What did we learn? 👉 PFS was improved with datopotamab 👉 OS data isn’t mature yet (we don’t know if patients live longer) 👉 Patients stayed on treatment longer (6.7 vs 4.1 months) 👉 Fewer dose reductions (20.8% vs 30.2%) 👉 Fewer interruptions (11.9% vs 24.5%) 👉 No difference in discontinuations The toxicities that stick out with this study are nausea, stomatitis, and ocular effects Half of patients experienced nausea and stomatitis, with 6.4% having grade 3+ stomatitis Patients had ophthalmologic assessments every 3 cycles and were advised to use artificial tears and avoid wearing contact lenses Table A4 in the study gives more details on what was included in stomatitis and ocular toxicities The package insert includes the use of cryotherapy during the infusion and QID eye drops/steroid mouthwash These were not mandated in the study and the publication doesn’t report how many patients used them so we’ll see what happens with real world use. --- I’m the Kelley in KelleyCPharmD 👋 and I help pharmacists learn the complex world of oncology

CAR-T therapies struggle to maintain cancer-killing ability without getting exhausted. This paper presents one gene edit to help fix that. The problem? The transcription factor FOXP3. It plays a key role in regulatory T cells, but is also transiently upregulated in effector T cells upon stimulation (limiting their function). Well. We can't have that. What the authors did: - Combined lentiviral transduction of CD19-CAR with non-viral CRISPR-Cas9 gene editing to knock out FOXP3 in human T cells. - Compared knocked-out CAR T to unmodified CAR T - Analyzed exhaustion markers, cytokine production, T cell phenotype, and epigenetic and transcriptomic changes post-editing. Results: - Knocked Oout CAR T cells showed enhanced cytokine production (IFNγ, TNFα, IL-2) and activation markers (CD154, CD137) upon repeated stimulation. - Despite improved function, exhaustion markers didn't increase in any significant way - And importantly, no major epigenetic disruptions or changes in exhaustion transcript levels were found after FOXP3 knockout. Sometimes, you don't need to add more things in. Just eliminate the stuff you don't need. Love it. Kudos to the authors - great job! Any thoughts on this approach? Drop them in the comments. Full paper link: https://lnkd.in/g84b-S4B

TOPO1i is today's favourite ADC payload mechanism, but what will be tomorrow's? The data is in, and it's decisive: the momentum behind Topoisomerase I inhibitor (TOPO1i) ADCs has accelerated into 2025. Is this justified, or are we seeing 'me too' developers jumping on the TOPO1i bandwagon? Let me know your thoughts in the comments. Market Validation 🟪 #Enhertu leads 2024 ADC sales at $3.8B, #Trodelvy at $1.3B. 🟪 All the $1B+ ADC licensing deals in 2024 (where payload mechanism disclosed) = TOPO1i ADCs. 🟪 Nearly 800 ongoing trials, and <3% clinical discontinuation rate. Innovation Pipeline 🟪 Faster clinical development timelines vs other payload classes. 🟪 72 bispecific TOPO1i ADCs in active development (4 in Phase 3). Include novel Nectin-4 x TROP-2 target pair. 🟪 While DXd continues to be the dominant payload, use of diverse scaffolds is indicative of innovation and opportunity for significant refinement within the Topo1i ADC class. 🟪 After failure of Merck Life Science's tubulin inhibitor ADC, Ladiratuzumab vedotin, TOPO1i ADCs are going after LIV-1. A target to watch, for sure. Competition Intensifies 🟪 5 of the top 10 TOPO1i developers are headquartered in China. Further 3 have headquarters in the Republic of Korea. The geographic diversification of expertise is accelerating platform evolution. 🟪 Last year, the FDA awarded Breakthrough Therapy Designation to sacituzumab tirumotecan, a second TOPO1i ADC using the TROP-2 targeting antibody, sacituzumab. Sacituzumab tirumotecan showed superior efficacy in pivotal Phase 2 TNBC trial, than seen with Trodelvy. Gilead Sciences may soon face serious competition. Key Takeaway: With proven commercial success, robust pipelines, and expanding global competition, this payload mechanism represents both immediate opportunity and long-term potential. What's your take on these developments? 🟪 If (when) approved in the US, will sacituzumab tirumotecan's superiority disrupt Trodelvy's $1.3B sales? 🟪 Will bispecific TOPO1i ADCs solve the resistance challenge? 🟪 TOPO1i is today's favourite payload mechanism, but what will be tomorrow's? Degraders? Immunostimulants? Analysis done using data from the Beacon ADC database. #antibodydrugconjugates #adc #drugdevelopment #pharmainnovation

Our lab's new paper (https://lnkd.in/gQa53Bqv) is now out at @NatureBiotech. We discovered that IL-10-expressing CAR T cells resist T cell dysfunction and mediate durable clearance of solid tumors and metastases (https://rdcu.be/duW2h) (1/) CAR T cells are remarkably effective in treating blood cancers, but not solid tumors. Antigen stimulation and immune suppressors cause T cell exhaustion and eventually dysfunction. Countering T cell exhaustion is required to enhance CAR T cell therapy for solid tumors. (2/) We previously reported that T cells upregulate IL-10-receptor expression when they become terminally exhausted (TCF1-TIM3+). IL-10–Fc fusion protein could reinvigorate terminally exhausted T cells and induce durable complete responses in solid tumors. https://lnkd.in/d7g34_i. (3/) However, to maintain adequate concentrations, multiple intratumoral injections of IL-10–Fc were necessary limiting the application. We designed and prepared IL-10-secreting CAR T using both mouse and human CAR T cells. (4/) We show that treatment with IL-10-CAR T cells leads to complete regression of established solid tumors and metastatic cancers across several cancer types in syngeneic and xenograft models, including colon cancer, breast cancer, melanoma, lymphoma, and pancreatic cancer. (5/) The superior antitumor effects of IL-10-producing CAR T cells challenge the conventional view that IL-10 is solely an immunosuppressive cytokine. Insights into the complex functions of cytokines could lead to further biomedical applications. (9/) The IL-10-secreting CAR T cell described here can potentially be a generalizable approach to prevent T cell exhaustion and metabolic dysfunction, which we term ‘metabolic armoring’. Extension to TCR-T, TIL, and other cell therapy can be expected. (10/) Clinical trials of IL-10-CD19-CAR T cells in patients with relapsed/refractory DLBCL or B-cell ALL are currently underway (NCT05715606, NCT05747157, NCT06120166). 12/12 patients have reached complete remission so far with 1-5% of typical CAR-T doses. (11/) We are thrilled that Research Briefing has highlighted our work (https://lnkd.in/g7UPWhB3). @Hongbo Chi, St Jude Children's Research Hospital comments “this is an interesting study with strong therapeutic relevance”. (12/) This work was led by Yang Zhao and Jiangqing Chen, and in close collaboration with Jie Sun lab Jie Sun, and @YugangGUO2 Lab, at Zhejiang University. We also thank the Santiago Carmona lab, and Pedro Romero lab, and other Li Tang lab members,  and core facilities - it was a real team effort! (13/) We are also grateful for the support from @EPFL and our funding sources @snf_ch, @krebsliga, @Krebs_Forschung, @ERC_Research, @XtalPi, #Kristian Gerhard Jebsen Foundation, #Anna Fuller Fund Grant, (14/)

What next for CAR-T Therapies? One of the most used cell therapies today are those based on CAR-T cells. Despite being extremely successful drugs leading to long term remission of several oncology diseases, there are still multiple lines of development ongoing, here are a few to consider in the space of rejuvenation or enhancement of these cells. 1. Checkpoint Inhibition: Using antibodies to block the PD-1/PD-L1 interaction can rejuvenate exhausted CAR-T cells. Blocking CTLA-4 to enhance T cell activation and proliferation. 2. Genetic Engineering: Using gene-editing tools, for example CRISPR, to knock out genes associated with T cell exhaustion, such as PD-1, LAG-3, or CTLA-4. Overexpressing genes that promote T cell survival and function, like IL-7R and c-myc. 3. Cytokine Support: Engineering CAR-T cells to secrete cytokines like IL-12, IL-15, or IL-18 to boost their activity and persistence. Alternatively administering cytokines or cytokine agonists externally to support CAR-T cell function. 4. Metabolic Reprogramming: Using drugs or genetic modifications to enhance CAR-T cell metabolism, improving their function and longevity. Mitochondrial biogenesis: Boosting mitochondrial function to sustain CAR-T cell energy demands. 5. Combination Therapies, small molecule inhibitors are useful in combining CAR-T cells with small molecule inhibitors that reverse exhaustion or enhance T cell activation. Immunomodulatory drugs,  using drugs like lenalidomide or pomalidomide to modulate the tumor microenvironment and support CAR-T cell activity. 6. Improved CAR Designs: Second-Generation CARs, designing CARs with co-stimulatory domains (e.g., CD28, 4-1BB) to provide additional activation signals. Third Generation by incorporating multiple co-stimulatory signals for enhanced activation and persistence. Armored CAR-T cells are engineering to express molecules like cytokines or enzymes that modify the tumor microenvironment to be more supportive of T cell activity. 7. Epigenetic Modulation: Using HDAC inhibitors or DNMT inhibitors to alter the epigenetic landscape of CAR-T cells, reversing exhaustion signatures. 8. Memory T Cell Engineering: Increasing Stem Cell Memory T Cells by engineering CAR-T cells to have a memory-like phenotype, supporting the longevity of active cells by enhancing their persistence and functionality. Promoting T Cell Differentiation, by encouraging a differentiation state that resists exhaustion by targeting T cell signaling pathways. 9. Tumor Microenvironment Modulation, targeting immunosuppressive cells by depleting or inhibiting regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), and other suppressive elements in the tumor microenvironment. By combining these strategies, we can significantly improve CAR-T cell performance and overcome the limitations posed by exhaustion.

SMRTS (Selective modRNA Translation System) form Lior Zangi lab. SMRTS is a dual-mRNA circuit that operates in an on/off manner, leveraging the unique, cell-specific expression of microRNAs. The first #mRNA carries a highly specific endoribonuclease (Cas6) together with a cell-specific microRNA recognition sequence on its 3’UTR. The second #mRNA carries the gene of interest along with a Cas6 recognition element. Its positive selection mechanism allows for targeted gene expression in desired cells while preventing expression in non-targeting cells with the microRNA. As a proof of concept, the paper demonstrated the cancer-specific variants, bcSMRTS and ccSMRTS, for breast and colon cancer, respectively. Systemic delivery of LNP-encapsulated SMRTS constructs yielded a 114-fold and 141-fold increase in tumor-specific expression in 4T1 and MC-38 models, respectively, while reducing off-target expression by over 380-fold. 

🔬 𝐍𝐞𝐱𝐭-𝐆𝐞𝐧𝐞𝐫𝐚𝐭𝐢𝐨𝐧 𝐀𝐃𝐂𝐬: 𝐃𝐨𝐮𝐛𝐥𝐢𝐧𝐠 𝐃𝐨𝐰𝐧 𝐨𝐧 𝐏𝐚𝐲𝐥𝐨𝐚𝐝𝐬 𝐢𝐧 𝐎𝐧𝐜𝐨𝐥𝐨𝐠𝐲 💥 The field of antibody–drug conjugates is undergoing a strategic evolution — and the spotlight is now on 𝐝𝐮𝐚𝐥-𝐩𝐚𝐲𝐥𝐨𝐚𝐝 𝐀𝐃𝐂𝐬, which aim to address the limitations of current single-warhead agents by 𝐜𝐨𝐦𝐛𝐢𝐧𝐢𝐧𝐠 𝐭𝐰𝐨 𝐝𝐢𝐬𝐭𝐢𝐧𝐜𝐭 𝐜𝐲𝐭𝐨𝐭𝐨𝐱𝐢𝐜 𝐦𝐞𝐜𝐡𝐚𝐧𝐢𝐬𝐦𝐬 in a single construct. 🚀 👉 These new ADCs are engineered to counteract tumor resistance, improve therapeutic efficacy, and expand the clinical reach across more difficult-to-treat cancers. The rationale: 𝐢𝐟 𝐨𝐧𝐞 𝐩𝐚𝐲𝐥𝐨𝐚𝐝 𝐩𝐚𝐭𝐡𝐰𝐚𝐲 𝐢𝐬 𝐛𝐥𝐨𝐜𝐤𝐞𝐝, 𝐭𝐡𝐞 𝐬𝐞𝐜𝐨𝐧𝐝 𝐦𝐚𝐲 𝐬𝐭𝐢𝐥𝐥 𝐛𝐞 𝐞𝐟𝐟𝐞𝐜𝐭𝐢𝐯𝐞 — increasing the chance of durable responses. 🧬 🧪 Early clinical entries like KH815 (Chengdu Kanghong Pharmaceutical Group Co.,Ltd.) and CLIO-8221 (Callio Therapeutics) exemplify divergent strategies: • KH815 leverages a 𝐓𝐎𝐏𝟏 𝐢𝐧𝐡𝐢𝐛𝐢𝐭𝐨𝐫 𝐚𝐧𝐝 𝐚𝐧 𝐑𝐍𝐀 𝐩𝐨𝐥𝐲𝐦𝐞𝐫𝐚𝐬𝐞 𝐈𝐈 𝐢𝐧𝐡𝐢𝐛𝐢𝐭𝐨𝐫, with staggered release linkers designed to hit both internal and peripheral tumor cells. • CLIO-8221 combines 𝐓𝐎𝐏𝟏 𝐚𝐧𝐝 𝐀𝐓𝐑 𝐢𝐧𝐡𝐢𝐛𝐢𝐭𝐨𝐫𝐬, targeting synthetic lethality in tumors with elevated DNA damage response activity. 📌 Payload pairing decisions are guided by synergy, tolerability, and strategic drug release kinetics. Companies like Sutro Biopharma, Inc., Araris Biotech AG, and CrossBridge are innovating around 𝐛𝐫𝐚𝐧𝐜𝐡𝐞𝐝 𝐥𝐢𝐧𝐤𝐞𝐫𝐬, 𝐡𝐲𝐝𝐫𝐨𝐩𝐡𝐢𝐥𝐢𝐜 𝐩𝐚𝐲𝐥𝐨𝐚𝐝 𝐦𝐚𝐬𝐤𝐢𝐧𝐠, and 𝐜𝐨𝐧𝐭𝐫𝐨𝐥𝐥𝐞𝐝 𝐛𝐲𝐬𝐭𝐚𝐧𝐝𝐞𝐫 𝐞𝐟𝐟𝐞𝐜𝐭𝐬 to fine-tune delivery and safety. 📉 𝐂𝐡𝐚𝐥𝐥𝐞𝐧𝐠𝐞𝐬 𝐫𝐞𝐦𝐚𝐢𝐧: • Balancing toxicity profiles across payloads • Fine-tuning drug-to-antibody ratios • Avoiding antagonistic effects between payloads • Demonstrating clinical synergy — not just additive benefit 🔄 With 𝐦𝐮𝐥𝐭𝐢𝐩𝐥𝐞 𝐜𝐥𝐢𝐧𝐢𝐜𝐚𝐥 𝐭𝐫𝐢𝐚𝐥𝐬 𝐧𝐨𝐰 𝐚𝐜𝐭𝐢𝐯𝐞 𝐚𝐧𝐝 𝐦𝐨𝐫𝐞 𝐢𝐧 𝐈𝐍𝐃-𝐞𝐧𝐚𝐛𝐥𝐢𝐧𝐠 𝐬𝐭𝐚𝐠𝐞𝐬, dual-payload ADCs represent a pivotal step in overcoming the resistance bottlenecks faced by traditional ADC therapies. 🎯 𝐊𝐞𝐲 𝐓𝐚𝐤𝐞-𝐀𝐰𝐚𝐲𝐬: • 🧬 Dual-payload ADCs combine two warheads to overcome resistance and expand efficacy. • 🔗 Linker technology and release kinetics are central to performance and safety. • 🧪 Preclinical data show promise, but human safety and tolerability remain key hurdles. • 🎯 Emerging pairings include TOP1 + ATR inhibitors, TOP1 + RNA Pol II inhibitors, and cytotoxins + immune agonists. 🔗 More information in a recent article by Asher Mullard on 𝑁𝑎𝑡𝑢𝑟𝑒 𝑅𝑒𝑣𝑖𝑒𝑤𝑠 𝐷𝑟𝑢𝑔 𝐷𝑖𝑠𝑐𝑜𝑣𝑒𝑟𝑦: https://lnkd.in/g4wcP4kT #AntibodyDrugConjugates #OncologyInnovation #DualPayloadADCs #CancerTherapeutics #DrugResistance #PrecisionMedicine #ClinicalDevelopment

Have your CAR T cells stopped running? I know, give 'em INO! In a paper published in Cancer Cell yesterday, scientists at Stanford found that supplementing CAR T cells with Inosine (INO) prevents their exhaustion, makes them more stem-like, and enhances their anti-tumor potency. This study provides yet another example of the pace at which this field is evolving and improving on this technology. ---- When T cells are exposed to antigen for too long, they become exhausted - this happens often within a tumor. When this happens, the T cells proliferation slows down and they can no longer mount an effective immune response. They also begin to express two proteins called CD39 and CD73. Unfortunately, CD39 and CD73 can metabolize the molecule ATP into Adenosine (Ado). This is not good because Ado is highly immunosuppressive and is known to inhibit T function. So, you get a scenario in which exhausted T cells, already struggling to mount an immune response, start making Ado, suppressing themselves even more. It's like when your parents grounded you when you were a kid so you ran to your room and slammed the door and got grounded for even longer - you're only making things worse. This negative feedback loop immunosuppressive snowball is a real problem when it comes to CAR T cells and scientists have been putting a lot of effort towards finding a solution. Well, in a study published in Cancer Cell yesterday, scientists at Stanford found that you can reverse the effects of Ado-mediated immunosuppression by overexpressing an enzyme called ADA-OE (Adenosine deaminase). ADA-OE overexpression in CAR T cells results in way LESS Ado and way MORE of a molecule called Inosine or IDO. They realized that simply giving the T cells IDO significantly improved their function by dramatically altering the cell's metabolism. 🔄🔃They found that INO reprogrammed the T cells metabolism by... 🚫 Inhibiting glycolysis ⬆ Increasing the cells reliance on oxidative phosphorylation (when the cell gets the majority of its energy from the mitochondria) - having healthy mitochondria is known to combat T cell exhaustion ♻ Increased glutaminolysis - this will enhance the flux of metabolites into and through the TCA cycle, increasing mitochondrial activity and suppressing exhaustion ⚙ Increasing the synthesis of polyamines - polyamines can influence the epigenetic landscape and activate transcriptional programs that confer stem cell-like characteristics - good for proliferation and self-renewal. When CAR T cells were grown in IDO-containing media they were able to significantly outperform non-IDO fed cells against a mouse model of leukemia. ----- 📫 📩 Check out my weekly newsletter (here: https://lnkd.in/eTQTkH9n) for more cancer research and cell therapy updates like this.

Intercellular mitochondrial transfer enhances T cell metabolic adaptability and anti-tumor efficacy   Adoptive T cell therapy (ACT) is an innovative personalized immunotherapy that aims to enhance the ability of the immune system to target and eliminate cancer cells by reintroducing patient T cells after they have been expanded or genetically modified in vitro. This approach has shown promising results in the treatment of hematological malignancies but faces challenges in solid tumors. A key obstacle is the suppressive tumor microenvironment (TME), which can impede T cell function by disrupting mitochondrial activity, leading to T cell exhaustion. This exhaustion is characterized by impaired mitochondrial function, transcriptional and epigenetic changes, which ultimately weaken the anti-tumor capacity of T cells and allow cancer cells to evade immune detection. Therefore, strategies to enhance T cell mitochondrial function and prevent exhaustion are essential for ACT studies against solid tumors. Traditional strategies to overcome T cell exhaustion include selecting T cell subtypes with robust mitochondrial function, using genetic engineering to enhance mitochondrial biogenesis, or adding antioxidants to maintain mitochondrial integrity during T cell expansion. However, these approaches are less effective when mitochondrial function or mitochondrial DNA (mtDNA) is already impaired. Recent studies have revealed the phenomenon of mitochondrial transfer between cells, reflecting the evolved role of mitochondria as symbionts. In particular, mitochondrial transfer via tunneling nanotubes (TNTs), membrane structures supported by F-actin, has been identified as a major transfer mechanism. Mitochondrial transfer can repair damaged cells, although cancer cells may also use this mechanism to acquire mitochondria from nearby immune or stromal cells to enhance their own growth. A recent study published in Cell by the laboratories of Luca Gattinoni at the Leibniz Institute for Immunotherapy and Shiladitya Sengupta at Harvard Medical School demonstrated that transfer of mitochondria from bone marrow stromal cells to T cells via TNTs significantly enhanced CD8+ T cell mitochondrial function. This mitochondrial enhancement improved T cell proliferation, infiltration, and efficacy in the TME, overcame T cell exhaustion, and significantly enhanced antitumor activity. This finding highlights the potential of mitochondrial transfer as a transformative strategy for cell therapy, providing a promising direction for advancing ACT in solid tumors. Reference [1] Jeremy Baldwin et al., Cell 2024 (DOI: 10.1016/j.cell.2024.08.029) #AdoptiveTCellTherapy #ACT #MitochondrialTransfer #CancerImmunotherapy #TCellExhaustion #TumorMicroenvironment #SolidTumorTherapy #CellTherapyInnovation #TunnelingNanotubes #CancerResearch