Bispecific antibodies targeting mutant RAS neoantigens

Jacqueline Douglass; Emily Han-Chung Hsiue; Brian J Mog; Michael S Hwang; Sarah R DiNapoli; Alexander H Pearlman; Michelle S Miller; Katharine M Wright; P Aitana Azurmendi; Qing Wang; Suman Paul; Annika Schaefer; Andrew D Skora; Marco Dal Molin; Maximilian F Konig; Qiang Liu; Evangeline Watson; Yana Li; Michael B Murphy; Drew M Pardoll; Chetan Bettegowda; Nickolas Papadopoulos; Sandra B Gabelli; Kenneth W Kinzler; Bert Vogelstein; Shibin Zhou

doi:10.1126/sciimmunol.abd5515

Bispecific antibodies targeting mutant RAS neoantigens

Sci Immunol. 2021 Mar 1;6(57):eabd5515. doi: 10.1126/sciimmunol.abd5515.

Authors

Jacqueline Douglass^{1

2

3}, Emily Han-Chung Hsiue^{1

2

3}, Brian J Mog^{1

2

3

4}, Michael S Hwang^{1

2

3}, Sarah R DiNapoli^{1

2

3}, Alexander H Pearlman^{1

2

3}, Michelle S Miller^{2

5

6}, Katharine M Wright^{2

5

6}, P Aitana Azurmendi^{2

5

6}, Qing Wang^{7

2

8}, Suman Paul^{1

2

3

9}, Annika Schaefer^{1

2

3}, Andrew D Skora^{1

2}, Marco Dal Molin^{1

10}, Maximilian F Konig^{1

2

3

11}, Qiang Liu^{1

2

3}, Evangeline Watson^{1

2

3}, Yana Li⁵, Michael B Murphy¹², Drew M Pardoll^{6

9}, Chetan Bettegowda^{1

3

13}, Nickolas Papadopoulos^{1

3

6

14}, Sandra B Gabelli^{5

9

15}, Kenneth W Kinzler^{1

3

6}, Bert Vogelstein^{7

2

3

6

14}, Shibin Zhou^{7

3

6}

Affiliations

¹ Ludwig Center, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
² Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.
³ Lustgarten Pancreatic Cancer Research Laboratory, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
⁴ Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
⁵ Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
⁶ Bloomberg~Kimmel Institute for Cancer Immunotherapy, Sidney Kimmel Comprehensive Cancer Center, Baltimore, MD 21287, USA.
⁷ Ludwig Center, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. [email protected] [email protected] [email protected].
⁸ Complete Omics Inc., Baltimore, MD 21227, USA.
⁹ Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
¹⁰ Department of Surgery, University of Maryland Medical Center, Baltimore, MD 21201, USA.
¹¹ Division of Rheumatology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21224, USA.
¹² Cytiva, Marlborough, MA 01752, USA.
¹³ Department of Neurosurgery, Johns Hopkins University School of Medicine, MD 21205, USA.
¹⁴ Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
¹⁵ Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Abstract

Mutations in the RAS oncogenes occur in multiple cancers, and ways to target these mutations has been the subject of intense research for decades. Most of these efforts are focused on conventional small-molecule drugs rather than antibody-based therapies because the RAS proteins are intracellular. Peptides derived from recurrent RAS mutations, G12V and Q61H/L/R, are presented on cancer cells in the context of two common human leukocyte antigen (HLA) alleles, HLA-A3 and HLA-A1, respectively. Using phage display, we isolated single-chain variable fragments (scFvs) specific for each of these mutant peptide-HLA complexes. The scFvs did not recognize the peptides derived from the wild-type form of RAS proteins or other related peptides. We then sought to develop an immunotherapeutic agent that was capable of killing cells presenting very low levels of these RAS-derived peptide-HLA complexes. Among many variations of bispecific antibodies tested, one particular format, the single-chain diabody (scDb), exhibited superior reactivity to cells expressing low levels of neoantigens. We converted the scFvs to this scDb format and demonstrated that they were capable of inducing T cell activation and killing of target cancer cells expressing endogenous levels of the mutant RAS proteins and cognate HLA alleles. CRISPR-mediated alterations of the HLA and RAS genes provided strong genetic evidence for the specificity of the scDbs. Thus, this approach could be applied to other common oncogenic mutations that are difficult to target by conventional means, allowing for more specific anticancer therapeutics.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Antibodies, Bispecific / immunology
Antibodies, Bispecific / pharmacology*
Antigens, Neoplasm* / chemistry
Antigens, Neoplasm* / immunology
Biomarkers, Tumor / antagonists & inhibitors*
Biomarkers, Tumor / chemistry
Biomarkers, Tumor / genetics
Biomarkers, Tumor / immunology
Cell Line
Cross Reactions
HLA Antigens / immunology
Humans
Lymphocyte Activation / genetics
Lymphocyte Activation / immunology
Mutant Proteins / antagonists & inhibitors*
Mutant Proteins / chemistry
Mutant Proteins / immunology
Mutation
Peptide Fragments
Protein Binding / immunology
T-Lymphocyte Subsets / immunology
T-Lymphocyte Subsets / metabolism
ras Proteins / antagonists & inhibitors*
ras Proteins / chemistry
ras Proteins / genetics
ras Proteins / immunology

Substances

Antibodies, Bispecific
Antigens, Neoplasm
Biomarkers, Tumor
HLA Antigens
Mutant Proteins
Peptide Fragments
ras Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding