Structure-Guided Engineering of Immunotherapies Targeting TRBC1 and TRBC2 in T Cell Malignancies

Ferrari, Mathieu; Baldan, Vania; Wawrzyniecka, Patrycja; Bulek, Anna; Kinna, Alex; Ma, Biao; Bughda, Reyisa; Akbar, Zulaikha; Srivast, Saket; Ghongane, Priyankha; Gannon, Isaac; Robson, Matthew; Sillibourne, James; Jha, Ram; Lim, Wen Chean; Hopkins, Jade R.; Welin, M.; Surade, Sachin; Dyson, Michael Eric; McCafferty, John; Cordoba, Shaun; Thomas, Simon; Logan, Derek D. T.; Sewell, Andrew K.; Maciocia, Paul; Onuoha, Shimobi; Pulé, Martin

doi:10.21203/rs.3.rs-1475171/v1

Cited by 5 publications

(13 citation statements)

References 27 publications

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“…We next asked whether the T cells identified in the EA-niches were in a functional or dysfunctional state. For this, we examined the ESGs in the EA-niche for the expression of the T-cell exhaustion signature 21 .As shown in Figure 7d , all the 5 EA-associated niches (EA-Q1 to EA Q5) showed the presence of the 10-gene T-cell exhaustion signature, including TRBC2 24 , LAG3 25 , HAVCR2 26 , and CSF1 27 .The number of spots expressing the T-cell exhaustion genes were significantly higher in the EA vs AA tumors ( Figure 7e ). Within each of the 5 EA-niches, the expression of each exhaustion gene is similarly higher in EA Figures 7 f-k).…”

Section: Resultsmentioning

confidence: 99%

Integrative spatial omics reveals distinct tumor-promoting multicellular niches and immunosuppressive mechanisms in African American and European American patients with TNBC

Zhu,

Balasubramanian,

Asirvatham

et al. 2024

Preprint

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Racial disparities in triple-negative breast cancer (TNBC) outcomes have been reported. However, the biological mechanisms underlying these disparities remain unclear. We integrated imaging mass cytometry and spatial transcriptomics, to characterize the tumor microenvironment (TME) of African American (AA) and European American (EA) patients with TNBC. The TME in AA patients was characterized by interactions between endothelial cells, macrophages, and mesenchymal-like cells, which were associated with poor patient survival. In contrast, the EA TNBC-associated niche is enriched in T-cells and neutrophils suggestive of an exhaustion and suppression of otherwise active T cell responses. Ligand-receptor and pathway analyses of race-associated niches found AA TNBC to be immune cold and hence immunotherapy resistant tumors, and EA TNBC as inflamed tumors that evolved a distinctive immunosuppressive mechanism. Our study revealed the presence of racially distinct tumor-promoting and immunosuppressive microenvironments in AA and EA patients with TNBC, which may explain the poor clinical outcomes.

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Section: Resultsmentioning

confidence: 99%

Integrative spatial omics reveals distinct tumor-promoting multicellular niches and immunosuppressive mechanisms in African American and European American patients with TNBC

Zhu,

Balasubramanian,

Asirvatham

et al. 2024

Preprint

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“…A mouse anti-human TRBC2 antibody was specifically designed and developed for diagnostic flow cytometry, following a previously described strategy. 14 In short, an anti-TRBC2 antibody was produced based on structural engineering and rational design of mutations on complementarity determining region (CDR)1 (T28K and Y32F) and CDR3 (A96N and N99M) of the JOVI.1 antibody (Kabat numbering scheme), resulting in switched antibody specificity from TRBC1 to TRBC2 ( Figure 1A ). The kinetic profile of the anti-TRBC2 antibody was studied against soluble TRBC1 + or TRBC2 + TCRs on a Biacore T200 surface plasmon resonance system (Cytiva, Marlborough, MA); and its thermal stability was assessed on a via Prometheus NT.48 nanoDSF differential scanning fluorimeter (NanoTemper, München, Germany) ( Supplemental Figure 1 ).…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…13 More recently, we employed computational biology and protein engineering to rationally design and produce mutant versions of JOVI.1 with switched specificity for TRBC2 and pre-clinical activity as CAR T-cell constructs targeting TRBC2 + T-cell malignancies. 14 The availability of complementary antibodies against TRBC1 and TRBC2 provides a unique opportunity to easily demonstrate T-cell clonality, in a fashion similar to detecting clonal B-cells based on kappa or lambda immunoglobulin light chain restriction. We hereby demonstrate the optimal performance of this approach for the confident laboratory diagnosis of T-cell neoplasms.…”

Section: Introductionmentioning

confidence: 99%

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Dual T-cell constant β chain (TRBC)1 and TRBC2 staining for the identification of T-cell neoplasms by flow cytometry

Horna,

Weybright,

Ferrari

et al. 2023

Preprint

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The diagnosis of leukemic T-cell malignancies is often challenging, due to overlapping features with reactive T-cells and limitations of currently available T-cell clonality assays. Recently developed therapeutic antibodies specific for the mutually exclusive T-cell receptor constant β chain (TRBC)1 and TRBC2 isoforms provide a unique opportunity to assess for TRBC-restriction as a surrogate of clonality in the flow cytometric analysis of T-cell neoplasms. To demonstrate the diagnostic utility of this approach, we studied 135 clinical specimens with (50) or without (85) T-cell neoplasia, in addition to 29 blood samples from healthy donors. Dual TRBC1 and TRBC2 expression was studied within a comprehensive T-cell panel, in a fashion similar to the routine evaluation of kappa and lambda immunoglobulin light chains for the detection of clonal B-cells. Polytypic TRBC expression was demonstrated on total, CD4+and CD8+T-cells from all healthy donors; and by intracellular staining on benign T-cell precursors. All neoplastic T-cells were TRBC-restricted, except for 5 cases (10%) lacking TRBC expression. T-cell clones of uncertain significance were identified in 15 samples without T-cell malignancy (13%), and accounted for smaller subsets than neoplastic clones (median: 4.7% vs. 73% of lymphocytes, p<0.0001). Single staining for TRBC1 produced spurious TRBC1-dim subsets in 21 clinical specimens (16%), all of which resolved with dual TRBC1/2 staining. Assessment of TRBC restriction by flow cytometry provides a rapid diagnostic method to detect clonal T-cells, and to accurately determine the targetable TRBC isoform expressed by T-cell malignancies.

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“…13 More recently, we employed computational biology and protein engineering to rationally design and produce mutant versions of JOVI.1 with switched speci city for TRBC2 and preclinical activity as CAR T-cell constructs targeting TRBC2 + T-cell malignancies. 14 The availability of complementary antibodies against TRBC1 and TRBC2 provides a unique opportunity to easily demonstrate T-cell clonality, in a fashion similar to detecting clonal B-cells based on kappa or lambda immunoglobulin light chain restriction. We hereby demonstrate the optimal performance of this approach for the con dent laboratory diagnosis of T-cell neoplasms.…”

Section: Introductionmentioning

confidence: 99%

Dual T-cell constant β chain (TRBC)1 and TRBC2 staining for the identification of T-cell neoplasms by flow cytometry

Horna,

Weybright,

Ferrari

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

The diagnosis of leukemic T-cell malignancies is often challenging, due to overlapping features with reactive T-cells and limitations of currently available T-cell clonality assays. Recently developed therapeutic antibodies specific for the mutually exclusive T-cell receptor constant β chain (TRBC)1 and TRBC2 isoforms provide a unique opportunity to assess for TRBC-restriction as a surrogate of clonality in the flow cytometric analysis of T-cell neoplasms. To demonstrate the diagnostic utility of this approach, we studied 135 clinical specimens with (50) or without (85) T-cell neoplasia, in addition to 29 blood samples from healthy donors. Dual TRBC1 and TRBC2 expression was studied within a comprehensive T-cell panel, in a fashion similar to the routine evaluation of kappa and lambda immunoglobulin light chains for the detection of clonal B-cells. Polytypic TRBC expression was demonstrated on total, CD4+ and CD8+ T-cells from all healthy donors; and by intracellular staining on benign T-cell precursors. All neoplastic T-cells were TRBC-restricted, except for 5 cases (10%) lacking TRBC expression. T-cell clones of uncertain significance were identified in 15 samples without T-cell malignancy (13%), and accounted for smaller subsets than neoplastic clones (median: 4.7% vs. 73% of lymphocytes, p < 0.0001). Single staining for TRBC1 produced spurious TRBC1-dim subsets in 21 clinical specimens (16%), all of which resolved with dual TRBC1/2 staining. Assessment of TRBC restriction by flow cytometry provides a rapid diagnostic method to detect clonal T-cells, and to accurately determine the targetable TRBC isoform expressed by T-cell malignancies.

show abstract

Structure-Guided Engineering of Immunotherapies Targeting TRBC1 and TRBC2 in T Cell Malignancies

Cited by 5 publications

References 27 publications

Integrative spatial omics reveals distinct tumor-promoting multicellular niches and immunosuppressive mechanisms in African American and European American patients with TNBC

Integrative spatial omics reveals distinct tumor-promoting multicellular niches and immunosuppressive mechanisms in African American and European American patients with TNBC

Dual T-cell constant β chain (TRBC)1 and TRBC2 staining for the identification of T-cell neoplasms by flow cytometry

Dual T-cell constant β chain (TRBC)1 and TRBC2 staining for the identification of T-cell neoplasms by flow cytometry

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