Innate Immune Response Following AAV Administration

Dauletbekov, Daniyar; Pfromm, Julia K.; Fritz, Allan K.; Fischer, Manuel

doi:10.1007/978-3-030-27378-1_27

Cited by 11 publications

(7 citation statements)

References 21 publications

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“…Further, the host cell's innate immune response typically serves as the initial barrier against viral dissemination. It has been documented that, within the conventional transfection parameters applied to the HEK293 cell line, the host cell's innate immune system tends to exhibit an upregulation 8,[26][27][28][29][30][31][32] . However, in our case, when comparing our optimal transfection conditions to the standard ones, we observed a notable downregulation or reduced expression of proteins associated with immune response (such as ASAH1, CREB1, CD81, POLR3F, STOM, ATP11B, ATP6V1C1, NFKB2) across the time points (Supplementary Data GO Analysis).…”

Section: Discussionmentioning

confidence: 99%

Temporal Insights into Molecular and Cellular Responses during rAAV Production in HEK293T Cells

Patra,

Tan,

Kok

et al. 2023

Preprint

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The gene therapy field is actively pursuing cost-effective, large-scale production of recombinant adeno-associated virus (rAAV) vectors for therapeutic applications, which demand high dosages. Enhanced yield is essential but presents technical and cost challenges. Strategies like suspension cell culture, transfection optimization, and cultivation conditions have shown moderate success but fall short for large-scale applications. In this study, we explore the host cell proteome of HEK293T cell lines used for AAV2 production under different transfection conditions (standard, sub-optimal, optimal) using SWATH-MS. To understand molecular and cellular mechanisms, we created a tailored spectral library for HEK293T-AAV interactions. Our gene ontology and pathway analysis revealed significant protein expression variations, particularly in processes related to cellular homeostasis, metabolic regulation, vesicular transport, ribosomal biogenesis, and cellular proliferation under optimal transfection conditions. These conditions increased rAAV titre by 50% compared to standard protocols. Furthermore, we identified alterations in host cell proteins crucial for AAV mRNA stability and gene translation, particularly regarding AAV capsid transcripts in optimal transfection conditions. This study provides insights into cellular mechanisms during rAAV production in HEK293T cells and offers potential advancements in scalability and cost-efficiency for gene therapy vector production.SignificanceGenerating AAV is challenging and the amount produced is limited, despite the success of gene therapy. Thus, it is necessary to enhance the productivity of host cells through various engineering techniques. The replication and transduction of viruses cause numerous modifications to the host cell’s proteome. Viruses exploit the processes of the host cell, influencing the availability, post-translational adjustments, associations, or placement of its proteins during the replication/growth cycle. Comprehending the fluctuations in biological processes during AAV replication is essential to grasping how virus-host relations shape the result of infection and viral protein expression.

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Section: Discussionmentioning

confidence: 99%

Temporal Insights into Molecular and Cellular Responses during rAAV Production in HEK293T Cells

Patra,

Tan,

Kok

et al. 2023

Preprint

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“…The initial innate immune responses are the first line of defense against viral vectors and help modulate subsequent adaptive immune responses ( 45 ). The gene enrichment in the innate immune response and immune effector process pathways discovered by WGCNA analysis demonstrates that the innate immune function of the spleen has been activated.…”

Section: Discussionmentioning

confidence: 99%

Integrated analysis of lncRNA-mediated ceRNA network involved in immune regulation in the spleen of Meishan piglets

Shi

Xu²,

Wu³

et al. 2022

Front. Vet. Sci.

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Meishan pigs are a famous local pig breed in China, with high fertility and early sexual maturity, and stronger immunity compared to other breeds. The spleen is the largest lymphoid organ in pigs and performs essential functions, such as those relating to immunity and haematopoiesis. The invasion of many pathogenic microorganisms in pigs is associated with spleen damage. Long non-coding RNAs participate in a broad range of biological processes and have been demonstrated to be associated with splenic immune regulation. However, the expression network of mRNAs and lncRNAs in the spleen of Meishan pigs remains unclear. This study collected spleen tissues from Meishan piglets at three different ages as a model, and mRNA and lncRNA transcripts were profiled for each sample. Additionally, 1,806 differential mRNAs and 319 differential lncRNAs were identified. A complicated interaction between mRNAs and lncRNAs was identified via WGCNA, demonstrating that lncRNAs are a crucial regulatory component in mRNA. The results show that the modules black and red have similar mRNA and lncRNA transcription patterns and are mainly involved in the process of the immune defense response. The core genes (DHX58 and IFIT1) and key lncRNAs (TCONS-00002102 and TCONS-00012474) of piglet spleen tissue were screened using the ceRNA network. The expression of these genes is related to the immune response of pigs. Our research may contribute to a further understanding of mRNA and lncRNA expression in the spleen of piglets, and provide new ideas to improve the disease resistance of piglets.

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“…In clinical trials involving AAV vectors, the used of corticosteroids has been a common approach to suppress the immune response [ 109 ]. Alternatively, more targeted approaches have been explored, such as modifying the capsids for immune evasion through techniques like site-directed mutagenesis, directed evolution, and utilizing exosomes.…”

Section: Aav Vector Therapymentioning

confidence: 99%

Innate Immune Response to Viral Vectors in Gene Therapy

Wang,

Shao

2023

Viruses

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Viral vectors play a pivotal role in the field of gene therapy, with several related drugs having already gained clinical approval from the EMA and FDA. However, numerous viral gene therapy vectors are currently undergoing pre-clinical research or participating in clinical trials. Despite advancements, the innate response remains a significant barrier impeding the clinical development of viral gene therapy. The innate immune response to viral gene therapy vectors and transgenes is still an important reason hindering its clinical development. Extensive studies have demonstrated that different DNA and RNA sensors can detect adenoviruses, adeno-associated viruses, and lentiviruses, thereby activating various innate immune pathways such as Toll-like receptor (TLR), cyclic GMP-AMP synthase–stimulator of interferon genes (cGAS-STING), and retinoic acid-inducible gene I–mitochondrial antiviral signaling protein (RLR-MAVS). This review focuses on elucidating the mechanisms underlying the innate immune response induced by three widely utilized viral vectors: adenovirus, adeno-associated virus, and lentivirus, as well as the strategies employed to circumvent innate immunity.

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Innate Immune Response Following AAV Administration

Cited by 11 publications

References 21 publications

Temporal Insights into Molecular and Cellular Responses during rAAV Production in HEK293T Cells

Temporal Insights into Molecular and Cellular Responses during rAAV Production in HEK293T Cells

Integrated analysis of lncRNA-mediated ceRNA network involved in immune regulation in the spleen of Meishan piglets

Innate Immune Response to Viral Vectors in Gene Therapy

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