# A next generation immune cell engineering platform to create multifunctional solid tumor cell therapies This page is a text version of a Portal Biotechnologies publication. The bibliographic record is labeled in the text so it can be read without the surrounding website. ## Bibliographic record - Title: A next generation immune cell engineering platform to create multifunctional solid tumor cell therapies - Document type: Presentation - Year: 2025 - Outlet: SITC Spring Scientific: Cellular Therapy for Solid Tumors - Cells: T cells - Cargos: mRNA, CRISPR RNPs, circRNA - Page URL: https://portal.bio/publications/2025-a-next-generation-immune-cell-engineering-platform-to-create-multifunctional-solid-tu--ajkGJBEAACwAc91d - Markdown URL: https://portal.bio/publications/2025-a-next-generation-immune-cell-engineering-platform-to-create-multifunctional-solid-tu--ajkGJBEAACwAc91d.md - Original file: https://portal-bio.cdn.prismic.io/portal-bio/aForFHfc4bHWiplC_SITC.03142025.pdf ## Publication text Cellular Therapy for Solid Tumors MARCH 12–14, 2025 | SAN DIEGO, CA & VIRTUAL A next generation immune cell engineering platform to create multifunctional solid tumor cell therapies Anil Narasimha, Ph.D Portal Bio March 14, 2025 Cellular Therapy for Solid Tumors | MARCH 12–14, 2025 | SAN DIEGO, CA & VIRTUAL Disclaimer: Employee of Portal Bio Cellular Therapy for Solid Tumors | MARCH 12–14, 2025 | SAN DIEGO, CA & VIRTUAL Portal Solves this Problem with a Simple, Scalable Approach Williams AR, Bao S, Miller DL. Biotechnol Bioeng. 1999 Hirsch, SM, et al. Keystone: Regulatory RNAs. 2023 Hirsch, SM, et al. Keystone: Emerging Cell Therapies. 2024 ### Mechanical Disruption Intracellular Deliverycells and Cargo Cells: Patient or donor-derived Cargo: RNA, CRISPR, protein, small mlcs… Cells Cargo For cell therapies, fundamental research, and bioproduction Portal’s Growth To DatePortal’s Growth To Date Early adopters across research and cell therapy including… Products Across Scales Research High Throughput Clinical 50+ Partners Armon Sharei, PhD (CEO): Founder & former CEO of SQZ Biotech. Invention to post-IPO with over $300M in equity financing and a + $1Bn Roche collaboration Alec Barclay (COO): Former COO of T2Biosystems. Sr. R&D leader at Becton Dickinson and Siemens. 5 FDA cleared / commercialized devices, 13 assays, 6 RUO/CE devices Anil Narasimha, PhD (CCO): Former CEO of Mekonos, nano-needles for precision cell delivery company. Raised over $30M in venture funding Current Transfection Technologies Have Many Limitations DNA mRNA, siRNA Proteins, small molecules, polymers, etc. ### Cell Types ### Cargo ### Lnp Virus ### Lnp Virus Electroporation Electroporation Model cell lines e.g., cancer cell lines Patient-derived cells e.g., T cells, NKs, iPSCs at scale Stewart and Sharei et al. Nature. 2016 doi: 10.1038/nature19764 Portal Excels Where Others Struggle Stewart and Sharei et al. Nature. 2016 doi: 10.1038/nature19764 DNA mRNA, siRNA Proteins, small molecules, polymers, etc. ### Cell Types ### Cargo Model cell lines e.g., cancer cell lines Patient-derived cells e.g., T cells, NKs, iPSCs at scale Portal enables cytosolic access for diverse cell types, across cargo materials Mechanical Delivery Has Limited Non-specific Impact on Normal T Cell Function Cells exposed to electroporation reveal high degree of misregulation Cells exposed to mechanoporation are more similar to controls misregulated genes (>2-fold change, p<0.05) 6 hrs 24 hrs misregulated genes (>2-fold change, p<0.05) Maintenance of Normal T cell Proliferation following Mechanoporation Ctrl ElectroMech 16x q threshold Electroporation vs. Control (6hr) Mechanical Delivery vs. Control (6hr) vs. Control (24hr) Mechanical Delivery vs. Control (24hr) 0.25 8,141 2,211 3,309 0 0.1 4,072 0 7 0 0.05 330 0 1 0 Table 1. FDR q-value thresholds for pairwise comparisons in unstimulated T cells DiTommaso T et al. 2018. PNAS; T cell data presented at ASGCT 2021 8 Product Lines Enable Seamless Transition Across Scales ### The Portal Solution “Powered by Portal” consumables compatible with common manufacturing equipment Available NOW for qualified early adopters GMP ready Q1 2025 Clinical Scale Integrations Proprietary instruments and consumables R&D Scale “Powered by Portal” consumables compatible with existing robotics Available NOW High Throughput Integrations Research-Scale Equipment Currently Available for Early Adopters Research-Scale System Gateway™ Consumable Delivery Cartridge MicroBooster™ Compact bench-top unit that controls delivery parameters easily fits in hood Cores with defined pore size for deformation 18mm 35mm Cell type-specific consumable for optimal mechanoporationmediated cargo delivery Robust Multiplex Delivery in Unstimulated T Cells ### Gfp Mixed dextran, mRNA, & RNP Hirsch, SM, et al. Keystone: Regulatory RNAs. 2023 Hirsch, SM, et al. Keystone: Emerging Cell Therapies. 2024 T cell Boost with CRISPR RNP, mRNA, and Dextran GFP Expressing, B2M Edited, and Dextran Delivered T Cells % GFP + and B2M - and Cas + Cells 85.7 Robust Sequential Delivery in Activated T Cells Activated sequential T cell boost with CRISPR RNP, mRNA Activate TRAC expression Control Boost ### Trac Rnp Activated T cells mRNA & B2M RNP 51% Viability of activated T cells B2M & TRAC Knockout & GFP Expressing T cells Control Boost TRAC expression Viability (%)Viability (%)% GFP + B2M - TRAC - T Cells T Cell Experiment: Demonstration of Cytotoxicity of mbIL2 and CAR circRNA Boosted T cells Activate with CD3/ CD28 Dynabeads for 48 hours Unstimulated T cells Activated T cells Cell Boost on D6 CD19 CAR cRNA mblL2-cRNA Activated T cells Engineered with Simultaneous circRNA Delivery for IL-2 and CD19 CAR TILs expressing mbIL2 and mbIL12 exhibit enhanced proliferation and function Implementing the Portal Solution: Clinical Scale Integrations ### The Portal Solution “Powered by Portal” consumables compatible with common manufacturing equipment Available NOW for qualified early adopters GMP ready in Q1 2025 Clinical Scale Integrations Proprietary instruments and consumables R&D Scale “Powered by Portal” consumables compatible with existing robotics Available in 2024 High Throughput Integrations ### Confidential Clinical Integration into Existing Systems Millibooster™ Cartridge ● Simple in-line integration with current GMP equipment ● Tube-weld or luer lock connection ● Capable of treating >1 Billion cells/min Clinical Scale Performance Protocol T Cells Boost Gateway Activation and Expansion Edited, Activated T Cells Lovo + Clinical Scale Booster RNA expressing and edited T Cells Robust cell expansion, viability, editing and mRNA expression Cell Expansion Viable Cell Count (M) 5 million/well at onset Viability Viability (%) B2M KO efficiency(%) mRNA Expression ### B2m Ko (%) Control Boost Control BoostD0 D4 D7 D10 D4 D7 D10 Fresenius Kabi – Point-of-Care CGT System © Copyright Fresenius Kabi AG - Enables same-day production of autologous, therapeutic cells at the patient point-of-care/bedside - Accepts WB or Apheresis starting material; multiple reagents and processing buffers - Contains accommodations for all manufacturing unit operations in a single closed system – Depletion and concentration – Washing and incubation – Cell selection and formulation – Cargo delivery - Supports clinical/commercial protocols by multiple sponsors *Supports patient-connected workflows in future releases ### In Development Chirgwin, D et al. 2021. ASGCT. Published Mechanical Delivery Data Demonstrates Robust Performance at Clinical Scales ### Application Opportunities Over 40 patients treated in SQZ clinical trials with mechanoporation systems Research-Scale Performance cGMP Performance ### Confidential 21 ### The Future of Cell Engineering Memory: mbIL-12 Eliminating Preconditioning: BCL-2 and mbIL-2 to enhance survival Killing: mbIL-12, mbIL-2, GZMB, Perforin Enhanced transduction: resting B cell transduction circRNA mRNA siRNA Multiplexed RNA-based Modifications to Enhance Cell Therapies Differentiation to target cell type (e.g. neurons) ### Confidential Broadening possibilities in research and therapeutics Now Beta Launched for Early Adopters! No limit on cell type Diverse range of cargos Proprietary Portal modifications coming soon! Cell Engineering Cell Analytics Up to 1B cells/min Scalable, cost-effective manufacturing Compatible with existing equipment Simple business model across scales ## Provenance Some publications reference SQZ Biotechnologies, a company Dr. Armon Sharei and colleagues previously founded. Portal is the next generation company leveraging Dr. Sharei's platform technology.