Granted Patent B2

US 12,195,723 · App. 17/093,011 · Granted Jan 14, 2025

Inventors: Feng Zhang (Cambridge, MA); Blake Lash (Cambridge, MA); Daniel Strebinger (Cambridge, MA)

Assignees: THE BROAD INSTITUTE, INC.; MASSACHUSETTS INSTITUTE OF TECHNOLOGY

C12N5/0635A61K39/4611A61K39/4612A61K39/4621A61K39/4622A61K39/4632A61K39/46433A61K39/464488A61K39/464491C07K14/70539C12Q1/6881A61K2239/31C12N2502/30C12N2510/00C12Q2600/158

Abstract

Claims (35)

1. An engineered B cell comprising:

a modified antigen presentation pathway as determined by detecting expression of an MHCI-reporter antigen complex and/or an MHCII-reporter antigen complex on the surface of the engineered B cell, wherein the modified antigen presentation pathway comprises one or more modified genes selected from LAMP2, PSMB8, TAP1, TAP2, ERAP1, CANX, CALR, TAPBP, PDIA3, CD74, HLA-DMA, HLA-DMB, HLA-DOA, TAPBPL, B2M, ERAP2, HLA-E, HLA-F, HLA-G, TLR4, TICAM1, LNPEP, TFEB, RAB27A, SEC61A1, RAB3A, RAB3B, BCAP31, EEA1, LAMP1, RAB4A, RAB5A, RAB11A, RAB7A, RHOB, PKN1, PIK3C3, TF, WASF1, DIAPH1, SYK, ICAM1, Cathepsin L, Cathepsin S, Cathepsin D, Cathepsin B, V-aTPase, NOX2, GILT, AAA, ATPase p97, Sec61, TAP, ERAP, IRAP, Sec22b, syntaxin 4, SNAP23, UNC93B1, MAPK, MAPKKK, MAPKK, p38, INK, ERK1/2, MMK3, MKK6, protein kinase C (PKC) alpha, PKC beta I, PKC beta II, atypical PKC ζ, PKC λ/ι, PKC δ, PKC ε, PKC θ, toll-like receptor (TLR)-1, TLR-2, TLR-4, TLR-5, TLR-7, and TLR-9, INF-gamma, IL-4, TNF-alpha, JAK, STAT1, STAT2, STAT3, RhoA, Cdc42, Rac1, Rho kinase; and

inducible or constitutive expression of one or more T-cell modulating agents selected from the group consisting of: CD27, CD28, CD40, CD122, CD137, OX40, GITR, ICOS CTLA4, PPP2CA, PPP2CB, PTPN6, PTPN22, PDCD1, ICOS (CD278), PDL1, KIR, LAG3, HAVCR2, BTLA, CD160, TIGIT, CD96, CRTAM, LAIR1, SIGLEC7, SIGLEC9, CD244 (2B4), TNFRSF10B, TNFRSF10A, CASP8, CASP10, CASP3, CASP6, CASP7, FADD, FAS, TGFBRII, TGFRBRI, SMAD2, SMAD3, SMAD4, SMAD10, SKI, SKIL, TGIF1, IL10RA, IL10RB, HMOX2, IL6R, IL6ST, EIF2AK4, CSK, PAG1, SIT1, FOXP3, PRDM1, BATF, VISTA, GUCY1A2, GUCY1A3, GUCY1B2, GUCY1B3, MT1, MT2, CD40, OX40, CD137, GITR, CD27, SUP-1 TIM, TNF, CD95L (FasL or Fas ligand), TRAIL, TL1A, and IL-2.

2. The engineered B cell of claim 1 , wherein the modified antigen presentation pathway is capable of processing and presenting an antigen such that the engineered B cell is capable of antigen-specific binding with a target T-cell and wherein the antigen is an extracellular antigen, an intracellular antigen produced by the engineered B cell, or both, wherein antigen-specific binding capability is measured by specifically detecting the MHC I-reporter antigen complex or the MHC II-reporter antigen complex on the engineered B cell.

3. The engineered B cell of claim 1 , wherein the modified antigen presentation pathway comprises one or more modifications in one or more cross-presentation pathway genes in the cross-presentation pathway such that the engineered B cell overexpresses one or more genes in the cross-presentation pathway, under expresses one or more genes in the cross-presentation pathway, lacks one or more genes in the cross-presentation pathway, or any combination thereof.

4. The engineered B cell of claim 1 , wherein the engineered B cell has increased MHC I extracellular antigen presentation relative to an unmodified antigen presenting cell.

5. The engineered B cell of claim 1 , wherein the engineered B cell is capable of being loaded with an extracellular antigen, is loaded with an extracellular antigen, or both, and wherein the extracellular antigen is optionally a peptide or polypeptide that optionally comprises an epitope tag.

6. The engineered B cell of claim 5 , wherein the engineered B cell expresses an engineered B cell receptor capable of specifically interacting with the epitope tag.

7. The engineered B cell of claim 2 , wherein the antigen is a target T-cell antigen.

8. The engineered B cell of claim 2 , wherein the target T-cell is a

a. CD8+ T-cell, CD4+ T-cell, CD25+ T-cell, or a combination thereof;

b. engineered T-cell, wherein the engineered T-cell is optionally a CAR T-cell;

c. non-pathogenic T-cell

d. pathogenic T-cell;

e. autoreactive T-cell; or

f. any combination thereof.

9. The engineered B cell of claim 8 , wherein the one or more T-cell modulating agents is/are capable of

a. eliminating or inhibiting one or more functions of the target T-cell and/or a cell within effective proximity of the target T-cell;

b. activating one or more functions of the target T-cell and/or a cell within effective proximity of the target T-cell; or

c. both (a) and (b).

10. The engineered B cell of claim 1 , wherein the one or more T cell modulating agents are immune checkpoint molecules, capable of inducing T-cell death, or both.

11. The engineered B cell of claim 1 , wherein the engineered B cell is capable of antigen-specific binding of a target T-cell, wherein antigen-specific binding capability is measured by specifically detecting an MHC I-antigen complex or an MHC II-antigen complex on the engineered B cell.

12. The engineered B cell of claim 1 , wherein the engineered B cell further comprises

a. a SynNotch receptor, a MESA receptor, or both and wherein expression, activation, or both of the one or more T-cell modulating agents is induced by activation of the SynNotch receptor, MESA receptor, or both;

b. a hemichannel capable of forming a gap junction between the engineered B cell and a hemichannel present on the target T-cell;

c. a trogocytic inducer, wherein the trogocytic inducer is capable of being expressed the surface of the engineered B cell, is optionally expressed on the surface of the engineered B cell, or both; or

d. a combination thereof.

13. The engineered B cell of claim 1 , wherein the engineered B cell is autologous.

14. The engineered B cell of claim 1 , wherein the engineered B cell is allogeneic.

15. The engineered B cell of claim 1 , wherein the engineered B cell is capable of self-inactivation or suicide.

16. A pharmaceutical formulation comprising:

an engineered B cell of claim 1 ; and

a pharmaceutically acceptable carrier.

17. The engineered B cell of claim 1 , wherein the modified antigen presentation pathway further comprises a modified MDY88 gene.

18. The engineered B cell of claim 1 , wherein the modified antigen presentation pathway further comprises a modified HLA-A gene, HLA-B gene, HLA-C gene, or any combination thereof.

Assignments (4)

Continuity (2)

References Cited (218)