Lack Of Immunogenicity Research Articles

Advances in veterinary ophthalmology: Acellular protein matrix from tilapia skin (Oreochromis Niloticus) in the recovery of corneal ulcers in dogs and cats

The acellular protein matrix from tilapia skin has excellent biomechanical properties, lack of immunogenicity and high biocompatibility in vitro and in vivo. Therefore, it provides an environment conducive to cell regeneration, specifically in more invasive and delicate applications, such as the recovery of corneal ulcers in dogs and cats. The objective of this literature review is to present the use of the scaffold as a new approach in veterinary ophthalmology in the treatment of corneal injuries in relation to conventional methods. When applied to corneal ulcers, the biomaterial acts as a tissue scaffold, facilitating cell migration, promoting angiogenesis and minimizing the inflammatory response; thus, significantly accelerating the healing process. It also improves corneal transparency, reduces recovery time and reduces post-treatment complications. Clinical trials involving dogs and cats have had significant success, demonstrating the applicability and effectiveness of this innovation in the context of veterinary ophthalmology. This study highlights both the effectiveness of the tilapia skin scaffold and emphasizes its potential to transform clinical practice by providing a sustainable, affordable and pioneering approach to treating corneal ulcers in small animals.

Clinical application and outcome of Tenon's patch graft: A comprehensive review.

Corneal perforations are potentially sight-threatening conditions if not promptly treated. Management depends on the size, location, cause, and severity of the perforation. Various methods, including tissue adhesives, amniotic membrane grafting (AMG), and keratoplasty, are available. However, the limited availability of donor tissue in emergencies poses a significant challenge. Recently, Tenon's patch grafting (TPG) has emerged as a promising treatment option due to its autologous nature, availability, and lack of immunogenicity. This review provides a comprehensive overview of various aspects of TPG, along with relevant images. A literature search using keywords such as "Tenon's patch graft," "Tenon's capsule," "corneal patch graft," "amniotic membrane," and "corneal perforations" yielded 37 articles, included in this review. TPG is suitable for cases involving corneal tissue degradation leading to perforation, impending perforation, or descemetocele, with lesion sizes ranging from 2 to 6 mm. For larger perforations, the procedure may be modified by combining it with AMG or tissue adhesives. Postoperatively, the graft undergoes characteristic changes, including epithelialization and collagen deposition, resulting in the formation of a thick scar. Complications such as delayed epithelialization, wound leaks, and graft dehiscence can occur. High success rates have been reported, ranging from 74% to 87%. However, TPG may result in suboptimal visual outcomes, particularly when the graft is centrally located, necessitating future keratoplasty for visual rehabilitation. Overall, TPG is a safe and cost-effective technique for restoring structural integrity in corneal thinning disorders and perforations, especially in emergency settings when donor tissue is unavailable.

Synthesis and Physicochemical Characterization of Gelatine-Based Biodegradable Aerogel-like Composites as Possible Scaffolds for Regenerative Medicine.

Regenerative medicine is an interdisciplinary field aiming at restoring pathologically damaged tissues and whole organs by cell transplantation in combination with proper supporting scaffolds. Gelatine-based ones are very attractive due to their biocompatibility, rapid biodegradability, and lack of immunogenicity. Gelatine-based composite hydrogels, containing strengthening agents to improve their modest mechanical properties, have been demonstrated to act as extracellular matrices (ECMs), thus playing a critical role in "organ manufacturing". Inspired by the lysyl oxidase (LO)-mediated process of crosslinking, which occurs in nature to reinforce collagen, we have recently developed a versatile protocol to crosslink gelatine B (Gel B) in the presence or absence of LO, using properly synthesized polystyrene- and polyacrylic-based copolymers containing the amine or aldehyde groups needed for crosslinking reactions. Here, following the developed protocol with slight modifications, we have successfully crosslinked Gel B in different conditions, obtaining eight out of nine compounds in high yield (57-99%). The determined crosslinking degree percentage (CP%) evidenced a high CP% for compounds obtained in presence of LO and using the styrenic amine-containing (CP5/DMAA) and acrylic aldehyde-containing (CPMA/DMAA) copolymers as crosslinking agents. ATR-FTIR analyses confirmed the chemical structure of all compounds, while optical microscopy demonstrated cavernous, crater-like, and labyrinth-like morphologies and cavities with a size in the range 15-261 µm. An apparent density in the range 0.10-0.45 g/cm3 confirmed the aerogel-like structure of most samples. Although the best biodegradation profile was observed for the sample obtained using 10% CP5/DMAA (M3), high swelling and absorption properties, high porosity, and good biodegradation profiles were also observed for samples obtained using the 5-10% CP5/DMAA (M4, 5, 6) and 20% CPMA/DMAA (M9) copolymers. Collectively, in this work of synthesis and physicochemical characterization, new aerogel-like composites have been developed and, based on their characteristics, which fit well within the requirements for TE, five candidates (M3, M4, M5, M6, and M9) suitable for future biological experiments on cell adhesion, infiltration and proliferation, to confirm their effective functioning, have been identified.

Dysregulation of Immune Tolerance to Autologous iPSCs and Their Differentiated Derivatives.

Induced pluripotent stem cells (iPSCs), capable of differentiating into any cell type, are a promising tool for solving the problem of donor organ shortage. In addition, reprogramming technology makes it possible to obtain a personalized, i.e., patient-specific, cell product transplantation of which should not cause problems related to histocompatibility of the transplanted tissues and organs. At the same time, inconsistent information about the main advantage of autologous iPSC-derivatives - lack of immunogenicity - still casts doubt on the possibility of using such cells beyond immunosuppressive therapy protocols. This review is devoted to immunogenic properties of the syngeneic and autologous iPSCs and their derivatives, as well as to the reasons for dysregulation of their immune tolerance.

First-in-man use of a cardiovascular cell-derived secretome in heart failure. Case report

BackgroundThere is increased evidence that the effects of stem cells can mostly be duplicated by administration of their secretome which might streamline the translation towards the clinics. MethodsThe 12-patient SECRET-HF phase 1 trial has thus been designed to determine the feasibility and safety of repeated intravenous injections of the extracellular vesicle (EV)-enriched secretome of cardiovascular progenitor cells differentiated from pluripotent stem cells in severely symptomatic patients with drug-refractory left ventricular (LV) dysfunction secondary to non-ischemic dilated cardiomyopathy. Here we report the case of the first treated patient (baseline NYHA class III; LV Ejection Fraction:25%) in whom a dose of 20x109 particles/kg was intravenously infused three times three weeks apart. FindingsIn addition to demonstrating the feasibility of producing a cardiac cell secretome compliant with Good Manufacturing Practice standards, this case documents the excellent tolerance of its repeated delivery, without any adverse events during or after infusions. Six months after the procedure, the patient is in NYHA Class II with improved echo parameters, a reduced daily need for diuretics (from 240 mg to 160 mg), no firing from the previously implanted automatic internal defibrillator and no alloimmunization against the drug product, thereby supporting its lack of immunogenicity. InterpretationThe rationale underlying the intravenous route is that the infused EV-enriched secretome may act by rewiring endogenous immune cells, both circulating and in peripheral organs, to take on a reparative phenotype. These EV-modified immune cells could then traffic to the heart to effect tissue repair, including mitigation of inflammation which is a hallmark of cardiac failure. FundingThis trial is funded by the French Ministry of Health (Programme Hospitalier de Recherche Clinique AOM19330) and the “France 2030” National Strategy Program (ANR-20-F2II-0003). It is sponsored by Assistance Publique-Hôpitaux de Paris.

Abstract 3197: Addressing drug metabolism and pharmacokinetics (DMPK) challenges of small molecule-drug conjugates (SMDCs)

Abstract Background: SMDCs are gaining momentum with potential advantages over antibody-drug conjugates (ADCs), eg, better tumor penetration and lack of immunogenicity. We have developed a novel platform technology with tailored solutions for ADCs and SMDCs with intracellular and extracellular cleavage options. Key features of the SMDC technology comprise of a small molecule ligand such as an αvβ3 integrin binder showing efficient tumor homing and a linker extracellularly cleaved by neutrophil elastase (NE) in the tumor microenvironment allowing for seamless release of different payload classes.1 Our lead SMDC VIP2362 is currently in a Phase 1 clinical trial for the treatment of advanced solid tumors (NCT05712889). SMDCs often have a shorter half-life compared with ADCs, which may result in more frequent dosing schedules. Here we show significant reduction of clearance and drastic prolongation of half-life of SMDCs achieved by adding a second binding moiety. Methods: Payloads such as kinesin spindle protein inhibitors (KSPi), cyclin-dependent kinase 9 inhibitors (CDK9i), and monomethyl auristatin E (MMAE) have been linked via elastase-cleavable linkers to one or, via a branched linker, to two αvβ3 integrin binding moieties. Cytotoxicity of SMDCs was measured in cell lines with or without NE. Monovalent and bivalent SMDCs were investigated in DMPK studies in rats while monitoring the conjugate and the released payload. Results: Payloads were coupled to the NE-cleavable peptides via different linkages. MMAE was conjugated via sterically hindered ester bonds, CDK9i was conjugated via sulfoximide bonds, and KSPi payloads were coupled via amide bonds. Each of these SMDCs was highly stable in plasma and buffer. Without NE, in vitro cytotoxicity of SMDCs in each cell line was poor with IC50 in the micromolar range. In contrast, in the presence of NE, potency increased, and cytotoxicity (IC50) was in the low nanomolar to picomolar range depending on the cell line and payload. In each case, the SMDCs reached similar potency as the free payload indicating efficient cleavage of the SMDC by NE. In DMPK studies of SMDCs in rat, none or trace levels of free payload (<0.1% as compared to the respective conjugates) was measured. With each of the SMDCs and independent of the conjugated payloads, a remarkable impact on exposure was observed when attaching a second binding ligand. The SMDCs with two ligands showed a 37- to 81-fold reduced plasma clearance as compared with the respective monovalent SMDCs, which parallels a 12- to 57-fold increase in half-life (eg, 2.1 h and 28 h for mono and bivalent MMAE SMDCs respectively). Initial in vivo studies in a cell line-derived xenograft model showed efficacy of bivalent SMDCs with once weekly dosing. Conclusion: Our technology platform comprises NE-cleavable SMDCs with promising options to tune the DMPK profile for a longer half-life. 1. Lerchen et al, AACR (2023). 2. Lerchen et al, Cancers (2023). Citation Format: Anne-Sophie Rebstock, Hans-Georg Lerchen, Harvey Wong, Beatrix Stelte-Ludwig, Mareike Wiedmann, Amy J. Johnson, Raquel Izumi, Ahmed Hamdy. Addressing drug metabolism and pharmacokinetics (DMPK) challenges of small molecule-drug conjugates (SMDCs) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3197.

Abstract 3210: Raptamer-drug conjugates as molecularly targeted cancer therapeutics

Abstract The use of aptamers as a therapeutic targeting moiety holds the promise of building on the success of antibody-drug conjugates (ADCs). While ADCs are an attractive modality, they face limitations, including limited penetration into solid tumors and a complicated production process for large-scale manufacturing. Aptamers have several potential advantages over antibodies as a targeting moiety including: (i) a smaller size that enables better tumor penetration; (ii) chemical synthesis which allows rapid scale-up and ease of incorporating modifications; (iii) a superior safety profile from lack of immunogenicity and (iv) room temperature stability. Therefore, the use of aptamers in conjunction with drugs (similar to ADCs) holds promise for development of targeted therapeutics. We have identified a novel therapeutic target for certain cancers (COF-01) using proteomics data and immunohistochemistry from patient-derived tumors. Both the COF-01 protein and related mRNA expression was shown to be significantly increased in non-small cell lung cancer, head and neck cancer and pancreatic cancer. Here, we present a new therapeutic approach that targets COF-01-positive cancer cells using next generation aptamer conjugates. We performed a cell- and protein-based selection using our proprietary bead-based combinatorial libraries to generate next-generation base-modified ssDNA aptamers (Raptamers) against COF-01. Binding affinities of the COF-01 Raptamer candidates were determined by concentration curves using biolayer interferometry. The dissociation constants for the COF-01 Raptamers ranged from 80 nM - 200 nM. COF-01-specific Raptamer-drug conjugates (Rap-DCs) were developed by conjugating the Raptamers to monomethyl auristatin E via a cleavable linker. In vitro efficacy data showed the COF-01 Rap-DC can selectively induce cytotoxicity in different types of COF-01-positive human cancer cells (A431, CAL 27, BxPC-3). There was a significant decrease in cell viability of COF-01-positive cells when compared to COF-01-negative control cells (IC50 values ranged from 5.19 nM - 14.11 nM). In vivo efficacy testing in a COF-01-postive cell-derived xenograft mouse model showed a significant decrease in tumor volume in animals treated with the COF-01 Rap-DC when compared to saline- and erlotinib-treated controls. This provides proof-of concept for the use of high-affinity Raptamers against novel molecular targets, such as COF-01. This platform can be used to develop Rap-DCs against emergent neo-antigens on cancer cells that have escaped chemotherapy. Citation Format: Stephanie P. Vega, Nancy E. Ward, Uksha Saini, Leniher Castan-Chibas, Garima Kaushik, Daniel Ciznadija, Dev Chatterjee, Michael J. Heffernan, Atul Varadhachary, Michael Ritchie. Raptamer-drug conjugates as molecularly targeted cancer therapeutics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3210.

Aptamers: Promising Reagents in Biomedicine Application.

Nucleic acid aptamers, often termed "chemical antibodies," are short, single-stranded DNA or RNA molecules, which are selected by SELEX. In addition to their high specificity and affinity comparable to traditional antibodies, aptamers have numerous unique advantages such as wider identification of targets, none or low batch-to-batch variations, versatile chemical modifications, rapid mass production, and lack of immunogenicity. These characteristics make aptamers a promising recognition probe for scientific research or even clinical application. Aptamer-functionalized nanomaterials are now emerged as a promising drug delivery system for various diseases with decreased side-effects and improved efficacy. In this review, the technological strategies for generating high-affinity and biostable aptamers are introduced. Moreover, the development of aptamers for their application in biomedicine including aptamer-based biosensors, aptamer-drug conjugates and aptamer functionalized nanomaterials is comprehensively summarized.

Hydrogels Based on Recombinant Spidroin Stimulate Proliferation and Migration of Human Corneal Cells.

The effect of recombinant spidroin (RS) hydrogel (HG) on anterior epithelial cells and keratocytes of the human cornea was studied in vitro. Corneal injuries are highly prevalent in developing countries according to the World Health Organization. Various technologies have recently been proposed to restore the damaged surface of the cornea. Use of biodegradable silk-based materials, including recombinant analogs of the spider silk protein spidroin, is an important avenue of research in the field of wound healing and corneal regeneration. Spidroins are well known for their optimal balance of strength and elasticity. Given their biological compatibility, lack of immunogenicity, and biodegradability, spidroins provide a biomaterial for tissue engineering and regenerative medicine. HGs based on RS rS2/12-RGDS were therefore tested for cytotoxicity toward isolated corneal epithelial cells and keratocytes with regard to possible changes in cell phenotype and migratory activity. A promising outlook and therapeutic potential were demonstrated for RS-based HGs.

Engineering U1-Based Tetracycline-Inducible Riboswitches to Control Gene Expression in Mammals.

Synthetic riboswitches are promising regulatory devices due to their small size, lack of immunogenicity, and ability to fine-tune gene expression in the absence of exogenous trans-acting factors. Based on a gene inhibitory system developed at our lab, termed U1snRNP interference (U1i), we developed tetracycline (TC)-inducible riboswitches that modulate mRNA polyadenylation through selective U1 snRNP recruitment. First, we engineered different TC-U1i riboswitches, which repress gene expression unless TC is added, leading to inductions of gene expression of 3-to-4-fold. Second, we developed a technique called Systematic Evolution of Riboswitches by Exponential Enrichment (SEREX), to isolate riboswitches with enhanced U1 snRNP binding capacity and activity, achieving inducibilities of up to 8-fold. Interestingly, by multiplexing riboswitches we increased inductions up to 37-fold. Finally, we demonstrated that U1i-based riboswitches are dose-dependent and reversible and can regulate the expression of reporter and endogenous genes in culture cells and mouse models, resulting in attractive systems for gene therapy applications. Our work probes SEREX as a much-needed technology for the in vitro identification of riboswitches capable of regulating gene expression in vivo.

Stem Cell-Based Therapy and Cell-Free Therapy as an Alternative Approach for Cardiac Regeneration.

The World Health Organization reports that cardiovascular diseases (CVDs) represent 32% of all global deaths. The ineffectiveness of conventional therapies in CVDs encourages the development of novel, minimally invasive therapeutic strategies for the healing and regeneration of damaged tissue. The self-renewal capacity, multilineage differentiation, lack of immunogenicity, and immunosuppressive properties of mesenchymal stem cells (MSCs) make them a promising option for CVDs. However, growing evidence suggests that myocardial regeneration occurs through paracrine factors and extracellular vesicle (EV) secretion, rather than through differentiation into cardiomyocytes. Research shows that stem cells secrete or surface-shed into their culture media various cytokines, chemokines, growth factors, anti-inflammatory factors, and EVs, which constitute an MSC-conditioned medium (MSC-CM) or the secretome. The use of MSC-CM enhances cardiac repair through resident heart cell differentiation, proliferation, scar mass reduction, a decrease in infarct wall thickness, and cardiac function improvement comparable to MSCs without their side effects. This review highlights the limitations and benefits of therapies based on stem cells and their secretome as an innovative treatment of CVDs.

Nerve regeneration using decellularized tissues: challenges and opportunities.

In tissue engineering, the decellularization of organs and tissues as a biological scaffold plays a critical role in the repair of neurodegenerative diseases. Various protocols for cell removal can distinguish the effects of treatment ability, tissue structure, and extracellular matrix (ECM) ability. Despite considerable progress in nerve regeneration and functional recovery, the slow regeneration and recovery potential of the central nervous system (CNS) remains a challenge. The success of neural tissue engineering is primarily influenced by composition, microstructure, and mechanical properties. The primary objective of restorative techniques is to guide existing axons properly toward the distal end of the damaged nerve and the target organs. However, due to the limitations of nerve autografts, researchers are seeking alternative methods with high therapeutic efficiency and without the limitations of autograft transplantation. Decellularization scaffolds, due to their lack of immunogenicity and the preservation of essential factors in the ECM and high angiogenic ability, provide a suitable three-dimensional (3D) substrate for the adhesion and growth of axons being repaired toward the target organs. This study focuses on mentioning the types of scaffolds used in nerve regeneration, and the methods of tissue decellularization, and specifically explores the use of decellularized nerve tissues (DNT) for nerve transplantation.

Selected aesthetic medicine treatment in patients with autoimmune diseases - a literature review


 
 
 Introduction Interest in cosmetic procedures has been increasing. Data on practices for correcting aesthetic deficits, including those caused by autoimmune diseases are limited. There are fears of disease reactivation and exacerbation of inflammation after surgery. Aim of the study The aim of the study was to investigate the potential influence and safety of aesthetic medicine treatment - hyaluronic acid, botulinum toxin and breast augmentation with implants in patients with autoimmune diseases. Materials and methods A search was conducted using PubMed and Google Scholar databases. Articles were searched in English using the following key words: autoimmune disease, immunogenicity, inflammation, connective tissue, aesthetic medicine. Results Hyaluronic acid is considered the best filler for cosmetic procedures, mainly due to its lack of immunogenicity. No clinical studies have identified contraindications to the administration of hyaluronic acid-based dermal fillers in patients with inflammatory diseases. Botulinum toxin is essential in alleviating involuntary functions currently associated with dystonia, spasticity, and autonomic disorders. The long-term safety of implants should be assessed concerning genetic and environmental factors and whether the autoimmune disease is in remission.Conclusion With the increasing number of aesthetic procedures using hyaluronic acid, there are more reports of potential adverse events. Most side effects are associated with hypersensitivity reactions. The use of hyaluronic acid products in patients with autoimmune diseases is controversial. However, no clinical studies have identified contraindications to the administration of hyaluronic acid-based fillers. Botulinum toxin is used for healing and pain control in patients with systemic sclerosis who have suffered phalangeal ulcers. Despite epidemiological studies conducted to assess the association between breast implants and autoimmune diseases the results remain inconclusive
 
 

Survivin‐Based Recombinant Overlapping Peptides Induce T Lymphocyte Cytotoxicity and Prolong the Survival in In Vivo Melanoma Model

AbstractAnti‐cancer vaccination emerged as a promising and cost‐effective immunotherapy, but the lack of immunogenicity has hindered the success of therapeutic vaccine development. To address this issue and improve therapeutic efficacy, this study presents the examination of recombinant overlapping peptides (ROP) based on the tumor‐associated antigen, survivin, on in vivo immunogenicity and anti‐tumor efficacy using a melanoma C57/BL mouse model. Results show that ROPs induce a remarkable 46.5% cytotoxic activity mediated by activated cytotoxic T lymphocytes, compared to only 3% in wild‐type (WT) survivin protein. Additionally, ROPs significantly reduce tumor size by over 500 mm3 and prolong survival rates in mice with zero deaths in the first 17 days and 30% survival at the end of day 23, while no mice immunized with WT survivin protein survive beyond day 20. ROPs combined with anti‐4‐1BB agonists lead to additional tumor size reduction by 500 mm3 and 70% survival on day 23. These findings underscore the importance of survivin as a trigger for tumor‐restricting immunity and provide therapeutic evidence of ROPs' anti‐tumor potential, especially when combined with other immunostimulants, such as anti‐4‐1BB agonists. ROPs and adjuvant immunostimulants represent a potent vaccine strategy for therapeutic purposes, increasing vaccine immunogenicity and improving survival against cancer.

Macrophages as Promising Carriers for Nanoparticle Delivery in Anticancer Therapy.

Macrophages play a critical role in the immune response due to their ability to recognize and remove pathogens, as well as present antigens, which are involved in inflammation, but they are also one of the most abundant immune cell populations present in the tumor microenvironment. In recent years, macrophages have become promising cellular carriers for drug and nanoparticle delivery to the tumor microenvironment, mainly due to their natural properties such as biocompatibility, degradability, lack of immunogenicity, long half-life in circulation, crossing biological barriers, and the possibility of migration and accumulation at a site of inflammation such as a tumor. For the effectiveness of this therapeutic strategy, known as "Trojan horse", it is important that the nanoparticles engulfed by macrophages do not affect their proper functioning. In our review, we discussed how the size, shape, chemical and mechanical properties of nanoparticles influence their internalization by macrophages. In addition, we described the promising research utilizing macrophages, their cell membranes and macrophage-derived exosomes as drug carriers in anticancer therapy. As a prospect of the wider use of this therapeutic strategy, we postulate its future application in boron delivery to the tumor environment in boron neutron capture therapy.

Novel Small Molecules in IBD: Current State and Future Perspectives.

Biologicals have dominated the therapeutic scenery in inflammatory bowel diseases (IBDs), namely ulcerative colitis (UC) and Crohn's disease (CD), for the past 20 years. The development of tofacitinib was the starting point for an era of small molecules after the era of biologicals. These new agents may challenge the use of biological agents in the future. They share properties that appeal to both patients and physicians. Low production costs, a lack of immunogenicity, and ease of use are only some of their benefits. On the other hand, patients and their physicians must manage the potential side effects of small molecules such as JAK inhibitors or S1P1R modulators. Here, we present agents that have already entered the clinical routine and those that are still being investigated in clinical trials.

Abstract 456: Raptamer-drug conjugates as molecularly targeted cancer therapeutics

Abstract While antibody-drug conjugates (ADC) are an attractive modality for targeted cancer therapy, they face limitations. These include a reduced volume of distribution with limited penetration into solid tumors, and an expensive, long and complicated production process, especially for large-scale manufacturing. These factors hinder the activity of these medications in certain tumors, and limit equitable access to all population segments. Use of aptamers as a therapeutic targeting moiety, holds the promise of building on the success of ADCs. Aptamers have several advantages over mAbs as a targeting moiety including: (i) smaller size that enables better tissue/tumor penetration; (ii) chemical synthesis which allows rapid and affordable scale-up; (iii) superior safety profile from lack of immunogenicity and short half-life which reduces off-target adverse events; and (iv) room temperature stability. Therefore, the use of aptamers in conjunction with drugs (similar to ADCs) holds promise for tumor management. We have recently identified a novel therapeutic target for certain cancers (COF-01) using analysis of proteomics data and immunohistochemistry from patient-derived tumors. COF-01 was shown to have highly specific increases in protein and mRNA expression in several cancers including non-small cell lung cancer, head and neck cancer and pancreatic cancer. COF-01 expression correlated with poor overall survival. Here, we present a new therapeutic approach that targets COF-01-positive cells using next generation drug conjugated aptamers. We have generated novel next-generation base-modified ssDNA aptamers (Raptamers) against COF-01 using a cell- and protein-based selection that utilizes our proprietary bead-based oligonucleotide libraries. The binding affinities of these COF-01 Raptamer candidates were determined by concentration curves using biolayer interferometry. Dissociation constants for the COF-01 Raptamers ranged from 80 nM - 200 nM. Anti-COF-01 Raptamer-drug conjugates (Rap-DCs) were developed by conjugating the Raptamers to monomethyl auristatin F via a cleavable linker. In vitro efficacy data has shown that the COF-01 Rap-DCs can selectively induce cytotoxicity in COF-01-positive human cells. There was a significant decrease in cell viability of COF-01-positive cells when compared to COF-01-negative control cells. This study provides proof-of concept for the use of high-affinity Raptamers against novel molecular targets, such as COF-01. This platform can be used to develop Raptamers against emergent neo-antigens on cancer cells that have escaped other therapies. Citation Format: Stephanie P. Vega, Dev Chatterjee, Atul Varadhachary, Michael Ritchie, Michael Heffernan, Daniel Ciznadija. Raptamer-drug conjugates as molecularly targeted cancer therapeutics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 456.

Non-Viral Carriers for Nucleic Acids Delivery: Fundamentals and Current Applications.

Over the past decades, non-viral DNA and RNA delivery systems have been intensively studied as an alternative to viral vectors. Despite the most significant advantage over viruses, such as the lack of immunogenicity and cytotoxicity, the widespread use of non-viral carriers in clinical practice is still limited due to the insufficient efficacy associated with the difficulties of overcoming extracellular and intracellular barriers. Overcoming barriers by non-viral carriers is facilitated by their chemical structure, surface charge, as well as developed modifications. Currently, there are many different forms of non-viral carriers for various applications. This review aimed to summarize recent developments based on the essential requirements for non-viral carriers for gene therapy.

Targeted Small Molecules for Systemic Lupus Erythematosus: Drugs in the Pipeline.

Despite the uncertainty of the pathogenesis of systemic lupus erythematosus, novel small molecules targeting specific intracellular mechanisms of immune cells are being developed to reverse the pathophysiological processes. These targeted molecules have the advantages of convenient administration, lower production costs, and the lack of immunogenicity. The Janus kinases, Bruton's tyrosine kinases, and spleen tyrosine kinases are important enzymes for activating downstream signals from various receptors on immune cells that include cytokines, growth factor, hormones, Fc, CD40, and B-cell receptors. Suppression of these kinases impairs cellular activation, differentiation, and survival, leading to diminished cytokine actions and autoantibody secretion. Intracellular protein degradation by immunoproteasomes, levered by the cereblon E3 ubiquitin ligase complex, is an essential process for the regulation of cellular functions and survival. Modulation of the immunoproteasomes and cereblon leads to depletion of long-lived plasma cells, reduced plasmablast differentiation, and production of autoantibodies and interferon-α. The sphingosine 1-phosphate/sphingosine 1-phosphate receptor-1 pathway is responsible for lymphocyte trafficking, regulatory T-cell/Th17 cell homeostasis, and vascular permeability. Sphingosine 1-phosphate receptor-1 modulators limit the trafficking of autoreactive lymphocytes across the blood-brain barrier, increase regulatory T-cell function, and decrease production of autoantibodies and type I interferons. This article summarizes the development of these targeted small molecules in the treatment of systemic lupus erythematosus, and the future prospect for precision medicine.

Safety and Biodistribution of Nanoligomers Targeting the SARS-CoV-2 Genome for the Treatment of COVID-19.

As the world braces to enter its fourth year of the coronavirus disease 2019 (COVID-19) pandemic, the need for accessible and effective antiviral therapeutics continues to be felt globally. The recent surge of Omicron variant cases has demonstrated that vaccination and prevention alone cannot quell the spread of highly transmissible variants. A safe and nontoxic therapeutic with an adaptable design to respond to the emergence of new variants is critical for transitioning to the treatment of COVID-19 as an endemic disease. Here, we present a novel compound, called SBCoV202, that specifically and tightly binds the translation initiation site of RNA-dependent RNA polymerase within the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome, inhibiting viral replication. SBCoV202 is a Nanoligomer, a molecule that includes peptide nucleic acid sequences capable of binding viral RNA with single-base-pair specificity to accurately target the viral genome. The compound has been shown to be safe and nontoxic in mice, with favorable biodistribution, and has shown efficacy against SARS-CoV-2 in vitro. Safety and biodistribution were assessed using three separate administration methods, namely, intranasal, intravenous, and intraperitoneal. Safety studies showed the Nanoligomer caused no outward distress, immunogenicity, or organ tissue damage, measured through observation of behavior and body weight, serum levels of cytokines, and histopathology of fixed tissue, respectively. SBCoV202 was evenly biodistributed throughout the body, with most tissues measuring Nanoligomer concentrations well above the compound KD of 3.37 nM. In addition to favorable availability to organs such as the lungs, lymph nodes, liver, and spleen, the compound circulated through the blood and was rapidly cleared through the renal and urinary systems. The favorable biodistribution and lack of immunogenicity and toxicity set Nanoligomers apart from other antisense therapies, while the adaptability of the nucleic acid sequence of Nanoligomers provides a defense against future emergence of drug resistance, making these molecules an attractive potential treatment for COVID-19.