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US 2024/0294942 A1Published application
Retargeted retroviral vectors and compositions or methods of use thereof
AssigneesThe Broad Institute, Inc., The General Hospital Corporation
InventorsKepler MEARS, Robert MANGUSO, Kathleen YATES
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Abstract
The invention features pseudotyped viral particles (e.g., lentiviral or gammaretroviral particles) and compositions and methods of use thereof, where the viral particles comprise a VHH domain.
Claims21
A pseudotyped viral particle comprising:
- (a) an envelope comprising a fusion protein, wherein the fusion protein comprises a viral envelope glycoprotein domain or fragment thereof fused to a single variable domain on a heavy chain (VHH) antibody (VHH) domain or antigen binding fragment thereof, wherein the VHH domain or antigen binding fragment thereof specifically binds an antigen present on a target cell;
- and
- (b) a heterologous polynucleotide.
A lentiviral particle comprising:
- (a) an envelope comprising a fusion protein, wherein the fusion protein comprises a VHH domain or antigen binding fragment thereof and a Morbillivirus hemagglutinin domain or fragment thereof, wherein the VHH specifically binds αMHCII;
- and
- (b) a polynucleotide encoding a guide polynucleotide and/or a Cas9 or another component of a genome editing system:
- or
- (a) an envelope comprising a fusion protein, wherein the fusion protein comprises a VHH domain or antigen binding fragment thereof and a Morbillivirus hemagglutinin domain or fragment thereof, wherein the VHH specifically binds αCD7, αCD8, or αCD4;
- and
- (b) a polynucleotide encoding a guide polynucleotide and/or a Cas9
4 more elements
A method for generating a pseudotyped viral particle for delivering a heterologous polynucleotide to a target cell, the method comprising:
- (a) displaying on the cell membrane of a eukaryotic cell a viral envelope glycoprotein domain or fragment thereof fused to a VHH domain or fragment thereof, wherein the VHH domain or fragment thereof specifically binds an antigen present on the target cell:
- (b) transfecting the eukaryotic cell with a viral transfer vector and one or more additional vectors encoding one or more viral polypeptides, thereby generating the pseudotyped viral particle for delivering a heterologous polynucleotide to the target cell.
A eukaryotic cell for generating a pseudotyped viral particle, the eukaryotic cell comprising:
- (a) a cell membrane comprising a viral envelope glycoprotein domain or fragment thereof fused to a VHH domain or fragment thereof, wherein the VHH domain or fragment thereof specifically binds an antigen present on a target cell:
- (b) a viral transfer vector;
- and
- (c) one or more additional vectors encoding one or more viral polypeptides.
1 more element
A mammalian expression vector comprising a polynucleotide encoding a polypeptide comprising a viral envelope glycoprotein domain or fragment thereof fused to a VHH domain or fragment thereof, wherein the VHH domain or fragment thereof specifically binds an antigen present on a target cell.
A fusion protein suitable for pseudotyping a viral particle, wherein the fusion protein comprises a viral envelope glycoprotein domain fused to a VHH domain, wherein the VHH domain or fragment thereof specifically binds an antigen present on a target cell, the fusion protein comprising a sequence with at least 85% sequence identity to a sequence selected from the group consisting of:
- MeV-Hwtc18-MHCII (N11)
- MGSRIVINREHLMIDRPYVLLAVLFVMFLSLIGLLAIAGIRLHRAAIYTAEIHKSLSTNLDVTN
- SIEHQVKDVLTPLFKIIGDEVGLRTPQRFTDLVKFISDKIKFLNPDREYDERDLTWCINPPERI
- KLDYDQYCADVAAEELMNALVNSTLLEARATNQFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYL
- SRGYNVSSIVTMTSQGMYGGTYLVGKPNLSSKGSELSQLSMHRVFEVGVIRNPGLGAPVFHMTN
- YFEQPVSNDFSNCMVALGELKFAALCHREDSITIPYQGSGKGVSFQLVKLGVWKSPTDMRSWVP
- LSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKNQALCENPEWAPL
- KDNRIPSYGVLSVNLSLTVELKIKIASGFGPLITHGSGMDLYKTNHNNVYWLTIPPMKNLALGV
- INTLEWIPRFKVSPALFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYD
- TSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDKKLWCRHFCVLADSESGGHI
- THSGMVGMGVSCTVTREDGTGGGGSGGGGSGGGGSAAAQVQLVQSGGGLVQPGGSLGLSCAASG
- NIGSRDNMGWYRQAPGKQREWVATISGYGIATYRDSVKGRFTVAKDTAKNIVSLQMNYLTTEDT
- AVYYCYAYAVDSRNIFWSQGTQVTVSGGGSGGGSYPYDVPDYA;
- MeV-Hwtc18-CD45 (32)
- MGSRIVINREHLMIDRPYVLLAVLFVMFLSLIGLLAIAGIRLHRAAIYTAEIHKSLSTNLDVTN
- SIEHQVKDVLTPLFKIIGDEVGLRTPQRFTDLVKFISDKIKFLNPDREYDERDLTWCINPPERI
- KLDYDQYCADVAAEELMNALVNSTLLEARATNQFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYL
- SRGYNVSSIVTMTSQGMYGGTYLVGKPNLSSKGSELSQLSMHRVFEVGVIRNPGLGAPVFHMTN
- YFEQPVSNDFSNCMVALGELKFAALCHREDSITIPYQGSGKGVSFQLVKLGVWKSPTDMRSWVP
- LSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKNQALCENPEWAPL
- KDNRIPSYGVLSVNLSLTVELKIKIASGFGPLITHGSGMDLYKTNHNNVYWLTIPPMKNLALGV
- INTLEWIPRFKVSPALFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYD
- TSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDKKLWCRHFCVLADSESGGHI
- THSGMVGMGVSCTVTREDGTGGGGGGGGSGGGGSAAAQVQLVQSGGGLVQPGGSLRLSCAASG
- RAFNSAAMGWYRQAPGSQRELVASISAGTASYADAVKGRFTISRDYAKNIIYLQMNSLKPDDTA
- VYFCNYRTTYTSGYSEDYWGQGTQVTVSGGGSGGGSYPYDVPDYA;
- MeV-Hwtc18-CD7 (humanized VHH10)
- MGSRIVINREHLMIDRPYVLLAVLFVMFLSLIGLLAIAGIRLHRAAIYTAEIHKSLSTNLDVTN
- SIEHQVKDVLTPLFKIIGDEVGLRTPQRFTDLVKFISDKIKFLNPDREYDERDLTWCINPPERI
- KLDYDQYCADVAAEELMNALVNSTLLEARATNQFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYL
- SRGYNVSSIVTMTSQGMYGGTYLVGKPNLSSKGSELSQLSMHRVFEVGVIRNPGLGAPVFHMTN
- YFEQPVSNDFSNCMVALGELKFAALCHREDSITIPYQGSGKGVSFQLVKLGVWKSPTDMRSWVP
- LSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKNQALCENPEWAPL
- KDNRIPSYGVLSVNLSLTVELKIKIASGFGPLITHGSGMDLYKTNHNNVYWLTIPPMKNLALGV
- INTLEWIPRFKVSPALFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYD
- TSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDKKLWCRHFCVLADSESGGHI
- THSGMVGMGVSCTVTREDGTGGGGSGGGGSGGGGSAAADVQLQESGGGSVQAGGSLRLSCAASG
- YTHSSYCMAWFRQAPGREREGVASIDSDGTTSYADSVKGRFTISQDNAKNTLYLQMNSLKPEDT
- AMYYCAARFGPMGCVDLSTLSFGHWGQGTQVTVSITGGGSGGGSYPYDVPDYA;
- MeV-Hwtc18-CD4 (03F11)
- MGSRIVINREHLMIDRPYVLLAVLFVMFLSLIGLLAIAGIRLHRAAIYTAEIHKSLSTNLDVTN
- SIEHQVKDVLTPLFKIIGDEVGLRTPQRFTDLVKFISDKIKFLNPDREYDFRDLTWCINPPERI
- KLDYDQYCADVAAEELMNALVNSTLLEARATNQFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYL
- SRGYNVSSIVTMTSQGMYGGTYLVGKPNLSSKGSELSQLSMHRVFEVGVIRNPGLGAPVFHMTN
- YFEQPVSNDFSNCMVALGELKFAALCHREDSITIPYQGSGKGVSFQLVKLGVWKSPTDMRSWVP
- LSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKNQALCENPEWAPL
- KDNRIPSYGVLSVNLSLTVELKIKIASGFGPLITHGSGMDLYKTNHNNVYWLTIPPMKNLALGV
- INTLEWIPRFKVSPALFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYD
- TSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDKKLWCRHFCVLADSESGGHI
- THSGMVGMGVSCTVTREDGTGGGGSGGGGSGGGGSAAAEVQLVESGGGSVQPGGSLTLSCGTSG
- RTFNVMGWFRQAPGKEREFVAAVRWSSTGIYYTQYADSVKSRFTISRDNAKNTVYLEMNSLKPE
- DTAVYYCAADTYNSNPARWDGYDFRGQGTQVTVSSGGGSGGGSYPYDVPDYA;
- and
- MeV-Hwtc18-CD8a (R3HCD27)
- MGSRIVINREHLMIDRPYVLLAVLFVMFLSLIGLLAIAGIRLHRAAIYTAEIHKSLSTNLDVTN
- SIEHQVKDVLTPLFKIIGDEVGLRTPQRFTDLVKFISDKIKFLNPDREYDERDLTWCINPPERI
- KLDYDQYCADVAAEELMNALVNSTLLEARATNQFLAVSKGNCSGPTTIRGQFSNMSLSLLDLYL
- SRGYNVSSIVTMTSQGMYGGTYLVGKPNLSSKGSELSQLSMHRVFEVGVIRNPGLGAPVFHMTN
- YFEQPVSNDFSNCMVALGELKFAALCHREDSITIPYQGSGKGVSFQLVKLGVWKSPTDMRSWVP
- LSTDDPVIDRLYLSSHRGVIADNQAKWAVPTTRTDDKLRMETCFQQACKGKNQALCENPEWAPL
- KDNRIPSYGVLSVNLSLTVELKIKIASGFGPLITHGSGMDLYKTNHNNVYWLTIPPMKNLALGV
- INTLEWIPRFKVSPALFTVPIKEAGEDCHAPTYLPAEVDGDVKLSSNLVILPGQDLQYVLATYD
- TSRVEHAVVYYVYSPSRSFSYFYPFRLPIKGVPIELQVECFTWDKKLWCRHFCVLADSESGGHI
- THSGMVGMGVSCTVTREDGTGGGGSGGGGSGGGGSAAAQVQLQESGGGSVQPGGSLRLSCAASG
- FTFDDYAMSWVRQVPGKGLEWVSTINWNGGSAEYAEPVKGRFTISRDNAKNTVYLQMNSLKLED
- TAVYYCAKDADLVWYNLSTGQGTQVTVSSAAAYPYDVPDYGSGGGSGGGSYPYDVPDYA.
63 more elements
Dependencies are inferred from the claim text.