CAR-T cell therapies are now the established treatments for patients with relapsed and/or refractory B cell lymphomas, B cell acute lymphoblastic leukemia, and multiple myeloma. Through deep depletion of targeted malignancy cells, CD 19-targeted and BCMA-target CAR-T treatment could lead to long-term remissions, in some cases curative. Inspired by the success of hematological cancers, CAR T cells are studied for diseases beyond oncology. A recent case study suggested deep depletion of B cells could alleviate the pathological progression and achieve sustained drug-free remission for autoimmune diseases such as systemic lupus erythematosus (SLE). Autoimmune diseases are “controlled” but incurable with the current treatment options due to their complex etiology. Cell therapies targeting autoreactive immune cells may offer a potentially curative approach in certain autoimmune diseases.

Key Learning Objectives:

• Review recent progress of cell therapies in autoimmune diseases
• Discuss the potential opportunities of CAR-T treatment in autoimmune disorders

Nanomaterials have already begun to play a pivotal role in overcoming the limitations of traditional drugs by enhancing drug delivery, improving bioavailability, overcoming biological barriers in the treatment and prevention of diseases. Spherical Nucleic Acids (SNAs) stand out as a versatile class of nanomaterials that have been utilized in applications spanning diagnostics and therapeutics. SNAs, comprised of a nanoparticle core densely functionalized with radially oriented oligonucleotides, offer unique advantages for nanomedicine. Their 3-dimensional structure renders SNAs with superior properties in comparison to linear nucleic acid molecules, including enhanced cellular uptake, resistance to nuclease degradation, and superior target affinity. These qualities make SNAs ideally suited for their application towards effective gene editing technologies. This presentation will cover two recent examples of employing SNAs in gene editing paradigms. First, an approach that enhances the therapeutic potential of small interfering RNAs (siRNAs) when formulated as SNAs is described. Until now, the gene silencing activity of SNAs had been hindered by endosomal entrapment. Here, a simple yet innovative strategy that employs calcium chloride (CaCl2) salting in SNA formulation results in enhanced cytosolic delivery and gene silencing activity by up to 20-fold across diverse cell lines, indicating a promising avenue for overcoming existing limitations. In a second, related story, SNAs are employed for the purposes of performing CRISPR/Cas9 systems for genome editing. In this work, Cas9 protein-cored SNAs, densely modified with DNA and preloaded with single-guide RNA, exhibit enhanced cellular uptake and stability, achieving notable genome editing efficiencies across multiple cell lines. The integration of GALA peptides and nuclear localization signals further enhances endosomal escape and nuclear localization. Taken together, this work underscores the importance of precise control over molecular interactions at the nanoscale and the potential of SNA-based nanotechnologies for gene editing applications. 

Learning Objectives:

• Spherical Nucleic Acids (SNAs), exhibit properties making them ideal constructs for nanomedicine and effective gene editing technologies. This talk describes routes of employing SNAs to markedly improve the cytosolic delivery of siRNAs and enhanced genome editing and delivery of CRISPR/Cas9 platforms.

There has been an explosion in the number of genetic medicines, including, cell & gene therapies in clinical development. These therapies contain thousands to billions more atoms than small molecules and biological drugs. Thus the design and manufacturing of these advanced therapies is much more complex than more established therapeutic modalities. GeneFab's synthetic biology team supports leading cell and gene therapy companies in designing their next-generation therapeutics. Our team has supported leading companies such as Senti Bio, Blue Rock Therapeutics, and Spark Therapeutics. Across these collaborations, GeneFab's scientists have designed custom CARs for NK cell therapy, master regulators for control of cell phenotype, small molecule regulated kill switches, and custom promoters for cell type and cell state-specific gene expression. We will present several case studies of how GeneFab's team has delivered on complex projects to enable our clients to develop their next-generation pipeline products.

Learning Objectives:

• GeneFab's synthetic biology team can support companies in the development of custom promoters, safety switches, and master regulators to enhance their cell and gene therapies for both safety and efficacy. 
• Designing custom CARs for NK cell therapy has enabled significant advancements in targeted therapies, showcasing innovative solutions for complex therapeutic challenges.

There’s an elephant in the room when it comes to cell therapy. Hundreds of thousands of patients are in need of these life saving therapies - but the vast majority can’t access the treatment. This is because cell therapy manufacturing is reliant on a highly-skilled and limited labor pool, which results in prohibitively high prices and lack of availability and affordability for most patients. 

Given approximately half of cell therapy manufacturing costs are driven by these labor challenges, bringing cell therapy scalability within reach requires cutting-edge automation. To meet this need, Multiply Labs launched a robotic system for cell therapy manufacturing that is proven to match the performance and reduce the costs of a manual process. This session from Fred Parietti, Ph.D., Co-founder and CEO of Multiply Labs will cover:

• How robotics radically improves the unit economics of cell therapy manufacturing. 
  • Estimates based on scientifically peer-reviewed industry data show that this technology can reduce the cost of typical GMP-level cell therapy manufacturing processes by 70%, on a per-product basis. 
• The power of automation to increase throughput and accelerate production timelines - for example, think of 18 products per incubator hosted in parallel, vs. one or two in a manual process. 
• How the robotic system overcomes potential regulatory and process barriers by operating market-leading GMP instruments that are already deployed to manufacture all currently approved cell therapies. This ensures manufacturers do not need to significantly change their existing processes. 
• Insight into the partnership between Multiply Labs and GenScript to automate the cell isolation phase of cell therapy manufacturing, reducing the complexity of this laborious process.
• The benefits of robotics when it comes to minimizing contamination risk and reducing human error. 
• How manufacturers can ensure they have the flexibility to automate any part of the process, while using less space to get the job done and ensuring the approach is future-proof.

It is critical that we expand patient access to these life-changing treatments. This session will explore the cutting-edge developments made this year in finally bringing automation to cell therapy manufacturing.

Tome Biosciences is leading the field of programmable genomic integration (PGI). Building on founding technologies obtained from MIT and Replace Therapeutics, Tome has developed and refined PGI resulting in stable integration of multiple kilobases of DNA at user-defined locations. Integrase-mediated PGI (I-PGI) uses a Cas9 nickase with a writing enzyme (reverse transcriptase or ligase) to write an integrase recognition site (‘beacon’) at a user-defined location in the genome. Beacon placement (BP) is followed by integrase-mediated integration of donor DNA. I-PGI enables therapeutic correction of genetic diseases through the insertion of functional genes at the endogenous location without dependency on a double-strand break (DSB) and is not dependent on homology-directed repair (HDR). This technology is agnostic to the size of the DNA template and, due to the orthogonal nature of the integrase: beacon system and lack of DSBs, this technology can be used to generate complex cell therapies via multiplex large gene insertion.

This presentation will focus on an overview of PGI and highlight the progress Tome has made in developing this technology into medicine for patients in need. We will present data showing efficient PGI in primary human cells (iPSCs, Primary Human Hepatocytes (PHH)), as well as our current progress in translating PGI into rodent and non-human primates (NHP).

Tome is developing this technology to treat a wide variety of both rare and common diseases. For in vivo gene therapy applications, PGI will enable the integration of a healthy copy of a gene at the endogenous location, meaning that a single product will work for the majority of the patient population, regardless of the specific mutation. For cell therapy applications, PGI enables the development of highly edited cell therapies, opening the door to a new generation of medicine specifically designed for common diseases such as autoimmunity.

Learning Objectives:
PGI is a new technology that combines site specificity of integrases with Cas9-based genome editing to allow large gene insertion/replacement at user-defined locations.

Immunotherapy using chimeric antigen receptor (CAR) expressing NK cells engineered via viral vectors has shown exceptional efficacy against hematologic cancers in clinical trials. This method, however, is limited in cargo capacity and large-scale manufacturing for clinical use is cost-prohibitive to most academic centers. We have addressed these shortcomings by engineering CAR-NK cells using non-viral transposon engineering that can be paired with concurrent CRISPR/Cas9 editing. We believe that this approach avails opportunities that will allow researchers to address some of the more difficult solid tumor types.

Here, we developed a CAR-NK therapy with specific advantages to killing mesothelin-positive ovarian cancers which tend to form solid tumors, a new area for CAR NK cells. We started by testing 3 different mesothelin binders named SS1, 121, and M912. Our transposon vector also encodes for secretion of the cytokine IL15 for increased persistence and function of CAR-NK cells. The SS1, 121, and M912 CAR NK cells with soluble IL15 (Armored-NK) were tested in vitro and in vivo. No CAR variant dominated during the in vitro testing, so all three proceeded into a pilot in vivo study. In this experiment, NSG mice were engrafted with a mesothelin high tumor cell line, A1847, engineered to express Luciferase. A1847 cells were injected 14 days prior to the Armored CAR NK injection. Both tumor cells and Armored CAR NK cells were delivered via intraperitoneal (IP) injection.

At week 6, control mice, 121 treated, and M912 treated mice had tumors beginning to grow exponentially while the SS1 CAR controlled tumor growth. This trend continued for 3 more weeks as the SS1-treated mice did not see significant tumor growth while all other mice had to be euthanized due to tumor burden. SS1 mice survived long term with the only detectable tumor being a mass that grew at the injection site, outside of the IP space. With these results, we plan to test additional edits and modifications to SS1 cytokine-armored CAR-NK cells before proceeding with a phase 1 clinical trial. We believe these edits will enable cytokine-armored CAR-NK cells to treat solid tumors.

Key Learning Objectives:
Describe natural killer (NK) cells offer an off-the-shelf therapy for cancer immunotherapy
Explain NK cells can be efficiently engineered using non-viral DNA transposons delivered as GenCircle plasmids
Explain transposon-engineered, cytokine-armored CAR-NK cells are effective against ovarian cancer in vitro and in vivo

The rapid advancements in mRNA-based therapeutics have ushered in a new era of medicine, promising revolutionary treatments for a wide range of diseases. However, despite the tremendous potential, several bottlenecks hinder the full realization of these therapies. This presentation will delve into the current challenges in mRNA therapeutics development, focusing on tissue distribution and the stability issues associated with traditional lipid nanoparticle (LNP) delivery systems. By leveraging novel mRNA delivery technologies and strong knowledge and expertise in mRNA medicines, we aim to enhance the stability, affordability, and efficacy of mRNA-based medicines and address unmet medical needs.

One of the primary hurdles in mRNA therapeutics is achieving targeted tissue distribution. The ability to deliver mRNA precisely to the desired cells and tissues is critical for therapeutic efficacy and minimizing off-target effects. Traditional LNPs, while effective in certain contexts, often face challenges in stability and specificity, limiting their widespread application. To address these issues, Zipcode Bio is pioneering the development of next-generation mRNA delivery systems. Our approach involves the use of novel nanoparticle formulations designed to enhance stability and cellular uptake. By modifying the composition and properties of these nanoparticles, we aim to improve their interaction with biological membranes and evade immune detection, thereby increasing their delivery efficiency to target tissues.

Our efforts also extend to the selection of therapeutic programs and diseases that can best leverage the advantages of targeted delivery and mRNA expression. By focusing on diseases where precise control of protein expression can yield significant therapeutic benefits, we aim to maximize the impact of our mRNA-based interventions. One example is to tackle prevalent diseases where traditional treatment options are limited or inadequate.

In summary, the development and optimization of mRNA delivery systems are critical to advancing mRNA-based medicines. Through innovative nanoparticle technology and strategic therapeutic targeting, Zipcode Bio is at the forefront of overcoming the current bottlenecks in mRNA therapeutics. This keynote will provide an exploration of these approaches, highlighting the progress made and the future directions for this promising field.

Learning Objectives:

• Understand the primary challenges in the development of mRNA-based therapeutics, including issues with tissue distribution and the stability of traditional lipid nanoparticle (LNP) delivery systems.
• Explore innovative approaches in mRNA delivery technology, focusing on the development of next-generation nanoparticles designed to enhance stability and organ specificity
• Identify the strategic selection of therapeutic programs and diseases that can benefit most from targeted mRNA delivery and precise protein expression to maximize therapeutic impact