Treating Tissue Factor–Positive Cancers with Antibody–Drug Conjugates That Do Not Affect Blood Clotting

Theunissen, Jan-Willem; Cai, Allen G.; Bhatti, Maryam M.; Cooper, Anthony B.; Avery, Andrew D; Dorfman, Ryan; Guelman, Sebastián; Levashova, Zoia; Migone, Thi‐Sau

doi:10.1158/1535-7163.mct-18-0471

Cited by 37 publications

(34 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…TF binds FVIIa and subsequentely triggers the downstream coagulation cascade leading to thrombin generation and fibrin-rich-clot formation. As we contributed to show [21], enhanced TF expression has been observed in several tumor cell lines and several types of human cancers [14,[22][23][24][25][26][27]. If TF also triggers cellular signaling events that facilitate tumor progression [12,28,29], a determinant role of TF-associated coagulation mechanisms in supporting metastasis has been demonstrated [10,12,17,30,31].…”

Section: Introductionmentioning

confidence: 67%

Vimentin prevents a miR-dependent negative regulation of tissue factor mRNA during epithelial–mesenchymal transitions and facilitates early metastasis

et al. 2020

View full text Add to dashboard Cite

Epithelial-mesenchymal transitions (EMTs) are high-profile in the field of circulating tumor cells (CTCs). EMT-shifted CTCs are considered to encompass pre-metastatic subpopulations though underlying molecular mechanisms remain elusive. Our previous work identified tissue factor (TF) as an EMT-induced gene providing tumor cells with coagulant properties and supporting metastatic colonization by CTCs. We here report that vimentin, the type III intermediate filament considered a canonical EMT marker, contributes to TF regulation and positively supports coagulant properties and early metastasis. Different evidence further pointed to a new post-transcriptional regulatory mechanism of TF mRNA by vimentin: (1) vimentin silencing accelerated TF mRNA decay after actinomycin D treatment, reflecting TF mRNA stabilization, (2) RNA immunoprecipitation revealed enriched levels of TF mRNA in vimentin immunoprecipitate, (3) TF 3′-UTR-luciferase reporter vector assays implicated the 3′-UTR of TF mRNA in vimentin-dependent TF regulation, and (4) using different TF 3′UTR-luciferase reporter vectors mutated for potential miR binding sites and specific Target Site Blockers identified a key miR binding site in vimentin-dependent TF mRNA regulation. All together, these data support a novel mechanism by which vimentin interferes with a miR-dependent negative regulation of TF mRNA, thereby promoting coagulant activity and early metastasis of vimentin-expressing CTCs.

show abstract

Section: Introductionmentioning

confidence: 67%

Vimentin prevents a miR-dependent negative regulation of tissue factor mRNA during epithelial–mesenchymal transitions and facilitates early metastasis

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Pig and rabbit chimeras of a human IgG1 antibody with a kappa light chain were created by grafting the variable regions of the heavy and light chains of a human anti–Tissue Factor antibody 25G1 [15] onto corresponding constant regions from pig and rabbit (S1 Table). While human and pig IgG1 is the predominant isotype in serum, rabbits only have one IgG isotype (one Cγ gene) [16, 17].…”

Section: Resultsmentioning

confidence: 99%

“…The extracellular domain (ECD) fragments of the FcγRIIIA receptors were synthesized and cloned into pcDNA3.1V5-HisA (ThermoFisher Scientific, V81020) with a C-terminal 6x Histidine tag (S2 Table). Expression and purification of the FcγRIIIA ECD fragments was accomplished using described methodology [15]. Proteins were buffer exchanged into PBS pH 7.4, and monomeric human and pig FcγRIIIA ECD fragments were isolated by preparative size exclusion chromatography (GE Healthcare Bio-Sciences, 28-9909-44).…”

Section: Methodsmentioning

confidence: 99%

Binding affinities of human IgG1 and chimerized pig and rabbit derivatives to human, pig and rabbit Fc gamma receptor IIIA

Bhatti¹,

Cai²,

Theunissen³

2019

PLoS ONE

Self Cite

View full text Add to dashboard Cite

While pigs and rabbits are used as models for human immune diseases, FcγR binding is poorly characterized in both test species. To evaluate antibody binding to FcγRIIIA, a receptor involved in antibody-dependent cellular cytotoxicity, chimerized antibodies were generated by grafting the variable regions of a human IgG1 onto scaffolds from both species. The affinities of the parent and chimeric antibodies to the FcγRIIIA proteins from all three species were determined. While the human IgG1 and rabbit IgG had similar affinities for each FcγRIIIA with notable differences across species, pig IgG1 only bound pig FcγRIIIA with appreciable affinity. Also, the functional pig and rabbit proteins described here can be used in future experiments, such as pharmacology and mechanism of action studies.

show abstract

“…In summary, accumulating preclinical and clinical evidence suggests that TF is a useful target for cancer therapy because of its common yet selective expression on several important tumor compartments and its minimal or restricted expression in normal cells. Several TF-targeting therapeutics, including ICON 35 , L-ICON 14 and CAR-NK (and -T) cells therapies and antibody-drug conjugates (ADC) 67,68 , are under preclinical and clinical evaluation. In particular, this study established the proof of concept of targeting TF as a new target for CAR-NK cellular immunotherapy for the treatment of TNBC and may warrant further investigation in TNBC patients.…”

Section: Discussionmentioning

confidence: 99%

Tissue factor as a new target for CAR-NK cell immunotherapy of triple-negative breast cancer

2020

Sci Rep

View full text Add to dashboard Cite

Triple-negative breast cancer (TNBC), representing ~15% of globally diagnosed breast cancer, is typically an incurable malignancy due to the lack of targetable surface targets for development of effective therapy. To address the unmet need for TNBC treatment, we recently determined that tissue factor (TF) is a useful surface target in 50-85% of patients with TNBC and developed a secondgeneration TF-targeting antibody-like immunoconjugate (called L-ICON) for preclinical treatment of TNBC. Using the chimeric antigen receptor (CAR) approach, here we develop and test TF-targeting CARengineered natural killer (TF-CAR-NK) cells that co-express CD16, the Fc receptor (FcγIII) to mediate antibody-dependent cellular toxicity (ADCC), for a preclinical assessment of immunotherapy of TNBC using TF-CAR-NK cell as single agent therapy and in combination with L-ICON. Our preclinical results demonstrate that TF-CAR-NK cells alone could kill TNBC cells and its efficacy was enhanced with L-ICON ADCC in vitro. Moreover, TF-CAR-NK cells were effective in vivo for the treatment of tnBc in cell lineand patient's tumor-derived xenograft mouse models. Thus, this study established the proof of concept of targeting TF as a new target in CAR-NK immunotherapy for effective treatment of TNBC and may warrant further preclinical study and potentially future investigation in TNBC patients.The adoptive transfer of chimeric antigen receptor (CAR)-expressing T and natural killer (NK) cells represents a novel cancer immunotherapy approach. The concept of the CAR is based upon the idea of expressing novel receptors on the T or NK cell surface that would enable the T and NK cell to identify corresponding antigens on the surface of a target cell. The basic CAR construct consists of an extracellular antigen-recognition domain, usually single-chain antibody variable fragments (scFv), attached to an extracellular spacer domain, a transmembrane domain of CD28 and a signaling cytoplasmic domain such as 4-1BB (CD137), OX40 (CD134), DAP10, ICOS and CD3zeta chain (CD3ζ). The most advanced application is the use of CAR-T cells targeting CD19, a surface antigen on B cell malignancies, which has demonstrated antitumor efficacy in patients with these cancers 1 . However, early-phase clinical trials of CAR T therapy also showed that this treatment is frequently associated with side effects 1 , some even causing life-threatening toxicity 2 , partly due to the fact that current CAR targets are expressed by both the malignant cells and normal cells, such as CD19, which is also expressed by normal B cells throughout the B cell lineage 1 . Also, one of the major challenges in CAR therapy is the heterogeneity of tumor antigens, for instance, antigen loss or low antigen density on the cancer cells 3 . To overcome these challenges, it would be ideal to develop CAR T or CAR NK cells that could target a surface molecule that is commonly yet selectively expressed by several major tumor compartments, including but not limited to, the cancer cell, cancer stem cell (CSC) and tum...

show abstract

Treating Tissue Factor–Positive Cancers with Antibody–Drug Conjugates That Do Not Affect Blood Clotting

Cited by 37 publications

References 51 publications

Vimentin prevents a miR-dependent negative regulation of tissue factor mRNA during epithelial–mesenchymal transitions and facilitates early metastasis

Vimentin prevents a miR-dependent negative regulation of tissue factor mRNA during epithelial–mesenchymal transitions and facilitates early metastasis

Binding affinities of human IgG1 and chimerized pig and rabbit derivatives to human, pig and rabbit Fc gamma receptor IIIA

Tissue factor as a new target for CAR-NK cell immunotherapy of triple-negative breast cancer

Contact Info

Product

Resources

About