Adeno-associated virus complex with improved expression of RUNX3 gene and uses for preventing or treating KRAS mutated lung cancer
Inventors
Bae, Suk Chul • Lee, You Soub • Chi, Xinzi • Lee, Ja Yeol
Assignees
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Publication Number
US-11767541-B1
Publication Date
2023-09-26
Expiration Date
Abstract
Provided is an adeno-associated virus (AAV) complex for expression of an RUNX3 gene including an asymmetrically modified inverted terminal repeat (ITR). The AAV complex has asymmetric ITRs in which one of the two ITRs is modified, thereby increasing self-replication efficiency in host cells and increasing expression efficiency of a delivered gene, and therefore, compared to existing AAV complexes, the AAV complex has an advantage of improved productivity and gene expression efficiency.
Core Innovation
The invention relates to an adeno-associated virus (AAV) complex comprising a polynucleotide sequence encoding a runt-related transcription factor 3 (RUNX3) protein positioned between a first inverted terminal repeat (ITR) and a second ITR. The first ITR is not modified, while the second ITR is modified, and the modification targets all or part of a stem-loop structure formed of a rep-binding element (RBE), RBE′, and regions A, A′, B, B′, C, C′, and D.
The disclosed AAV complex uses asymmetrically modified ITRs, where only one of the two ITRs is modified. The modification alters the second ITR stem-loop structure, including insertion, deletion, substitution, or preventing hairpin formation, and includes examples where defined regions after RBE are deleted to produce a modified second ITR.
The invention addresses limited productivity and gene expression/productivity associated with unmodified or symmetrically modified ITRs. The disclosure states that the asymmetry increases AAV self-replication efficiency and improves gene expression/productivity relative to unmodified or symmetrically modified ITR controls.
The document further describes use for preventing or treating KRAS-mutated lung cancer with cancer-cell selectivity, with enhanced apoptosis/necrosis effects in KRAS-mutated lung cancer models and limited effects in normal lung epithelial cells.
Claims Coverage
The independent claim covers an AAV complex with RUNX3 expression between first and second ITRs, where the first ITR is unmodified and the second ITR is asymmetrically modified by altering the stem-loop structure defined by RBE/RBE′ and named regions A, A′, B, B′, C, C′, and D. Dependent claims add refinements to the genome design and specify KRAS-mutated lung cancer, including formulation aspects and optional constraints such as AAV serotype selection.
Asymmetrically modified ITRs for RUNX3 expression in an AAV complex
An AAV complex comprising a polynucleotide sequence encoding a RUNX3 protein between a first ITR and a second ITR, wherein the first ITR is not modified and the second ITR is modified, and wherein the second ITR all or part of a stem-loop structure formed of a rep-binding element (RBE), RBE′, A, A′, B, B′, C, C′, and D regions is modified.
Second ITR stem-loop modification via insertion, deletion, or substitution
The AAV complex wherein the second ITR stem-loop structure is modified by insertion, deletion, or substitution.
Deletion-based stem-loop modification of defined regions in the second ITR
The AAV complex wherein the second ITR, including all or part of a stem-loop structure formed of RBE, RBE′, and regions A, A′, B, B′, C, C′, and D, is deleted.
Treatment of KRAS-mutated lung cancer using the AAV complex
Treating KRAS-mutated lung cancer by administering an effective amount of the AAV complex to a subject in need thereof.
Pharmaceutical composition for preventing or treating KRAS-mutated lung cancer
A pharmaceutical composition for preventing or treating KRAS-mutated lung cancer comprising the AAV complex and a pharmaceutically acceptable carrier.
Overall, the claim set centers on an AAV genome design with RUNX3 between an unmodified first ITR and an asymmetrically modified second ITR, where the second ITR stem-loop structure defined by RBE/RBE′ and regions A, A′, B, B′, C, C′, and D is modified, including insertion, deletion, substitution, or deletion of the stem-loop structure. Dependent claims further define KRAS-mutated lung cancer treatment and a corresponding pharmaceutical composition.
Stated Advantages
Increases AAV self-replication efficiency.
Improves gene expression/productivity versus unmodified or symmetrically modified ITR controls.
Provides cancer-cell selectivity for preventing or treating KRAS-mutated lung cancer.
Enhances apoptosis/necrosis in KRAS-mutated lung cancer models with limited effects in normal lung epithelial cells.
Documented Applications
Preventing or treating KRAS-mutated lung cancer using an AAV complex encoding RUNX3 with asymmetrically modified ITRs.
Treatment application is described for non-small cell lung cancer and small cell lung cancer contexts, including KRAS-mutated lung cancer models and xenograft models.
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