Recombinant herpes simplex virus having expression cassette expressing fused protein of cancer cell-targeting domain and extracellular domain of HVEM and use thereof
Inventors
KWON, Heechung • Baek, Hyunjung • JOO, Hyun Yoo
Assignees
Korea Institute of Radiological and Medical Sciences • Gencellmed Inc
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Publication Number
US-11421017-B2
Publication Date
2022-08-23
Expiration Date
Abstract
The present invention relates to a recombinant herpes simplex virus (HSV) containing an expression cassette capable of expressing a fused protein of a cancer-cell-targeting domain and an extracellular domain of HVEM and the use thereof. When the recombinant HSV infects and enters target cells, which are cancer cells, HSV proliferates, and an adapter, which is the fused protein, is expressed in the cells and is released to the outside of the cells along with the proliferated HSV virion upon cell lysis, or is released even before the virion is released due to cell lysis when the adapter contains a leader sequence, and the fused protein released to the outside of the cells acts to induce the HSV virion to infect surrounding cancer cells expressing a target molecule recognized by the cancer-cell-targeting domain or to increase the infection efficiency thereof.
Core Innovation
The invention relates to a recombinant herpes simplex virus (HSV) engineered with an adapter expression cassette that expresses a fusion protein comprising a cancer-cell-targeting domain and an extracellular domain of HVEM. The adapter expression cassette is inserted into the genome of the herpes simplex virus at specified loci without inhibiting proliferation of the herpes simplex virus.
The recombinant HSV further includes mutation(s) in the gene encoding glycoprotein D (gD) so that the encoded gD is prevented from binding to nectin-1 while still retaining the ability of gD to bind to HVEM. This design is described as enabling infection via HVEM while restricting entry that depends on nectin-1 binding, in combination with the adapter fusion protein that includes the HVEM extracellular domain.
The disclosed system includes multiple defined insertion loci in the HSV genome for the adapter expression cassette, including between UL3 and UL4, between UL26 and UL27, between UL37 and UL38, between UL48 and UL49, between UL53 and UL54, or between US1 and US2. Dependent disclosures refine the adapter fusion protein by specifying HVEM extracellular domain variants and defining a linkage between the cancer-cell-targeting domain and the extracellular domain of HVEM through a linker peptide.
Claims Coverage
The partial content provides one independent claim, supported by multiple dependent claims that define specific fusion components, linker features, targeting specificity, and an optional second expression cassette payload, while maintaining the overall HVEM-adapter plus gD mutation framework. Across the claims presented, the inventive features focus on genome insertion of an adapter expression cassette at defined HSV loci and gD mutation to prevent nectin-1 binding while retaining HVEM binding, with dependent features specifying fusion ordering and payload options.
HVEM extracellular domain adapter cassette inserted at defined HSV loci
A recombinant herpes simplex virus (HSV) includes an adapter expression cassette expressing a fusion protein of a cancer-cell-targeting domain and an extracellular domain of HVEM inserted into a genome of the herpes simplex virus without inhibiting proliferation, wherein the adapter expression cassette is inserted between UL3 and UL4, between UL26 and UL27, between UL37 and UL38, between UL48 and UL49, between UL53 and UL54 or between US1 and US2.
gD mutated to prevent nectin-1 binding while retaining HVEM binding
In the recombinant HSV, the gene encoding glycoprotein D (gD) is mutated so as to prevent the encoded gD from binding to nectin-1 while still retaining the ability of said gD to bind to HVEM.
Cancer-cell targeting domain and HVEM extracellular domain fused via a defined linker range
The cancer-cell-targeting domain and the extracellular domain of HVEM are linked via a linker peptide comprising 1 to 30 amino acids.
HER2 scFv targeting moiety with defined VH and VL order
The cancer-cell-targeting domain recognizes and binds HER2 and is an scFv formed by linking VH (SEQ ID NO: 1) and VL (SEQ ID NO: 2) through a linker peptide in the order VH–linker–VL.
Selected HVEM extracellular domain variants
The extracellular domain of HVEM is specified as one of HveA82, HveA87, HveA102, or HveA107, each defined by particular amino acid sequences (SEQ ID NOs referenced).
Second expression cassette for immune-modulating and tumor-targeting genes inserted without inhibiting proliferation
A second expression cassette encoding one of several selected cytokine/chemokine/immune-checkpoint/co-stimulatory/TGFβ-antagonist/heparinase/VEGFR2-antagonist/prodrug-activating genes is further inserted into the viral genome without inhibiting herpes simplex virus proliferation.
Within the provided claim set, coverage centers on a recombinant HSV that couples a cancer-cell-targeting domain to an HVEM extracellular domain via an adapter expression cassette inserted at specified HSV loci without inhibiting HSV proliferation, together with gD mutations that prevent nectin-1 binding while retaining HVEM binding. Dependent claims further narrow the HVEM extracellular domain variants, the linker peptide range, the structural organization of HER2 scFv components, and optionally add a second payload expression cassette for selected immune-modulating or tumor-targeting genes without inhibiting HSV proliferation.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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