Retargeting of T cells to prostate stem cell antigen expressing tumor cells: Comparison of different antibody formats

Feldmann, Anja; Stamova, Slava; Bippes, Claudia C.; Bartsch, Holger; Wehner, Rebekka; Schmitz, Marc; Temme, Achim; Cartellieri, Marc; Bachmann, Michael

doi:10.1002/pros.21315

Cited by 62 publications

(124 citation statements)

References 29 publications

Supporting

Mentioning

120

Contrasting

Order By: Relevance

“…47 Purification of the his-tagged recombinant proteins from cell culture supernatant was performed by affinity chromatography on Ni-NTA columns (Qiagen, Hilden, Germany) according to previously published protocols. [44][45][46][47] For prokaryotic expression of the a-EGFR TM the E. coli strain SHuffle Ò T7 (New England BioLabs GmbH, Frankfurt am Main, Germany) was transformed with the vector pET28a_a-EGFR TM using standard heat shock method. One single clone of genetically modified bacteria was grown over night C, 10,000g), the EGFR-specific TM was purified by Ni-NTA affinity chromatography.…”

Section: Expression and Purification Of Recombinant Antibodiesmentioning

confidence: 99%

See 1 more Smart Citation

A novel nanobody-based target module for retargeting of T lymphocytes to EGFR-expressing cancer cells via the modular UniCAR platform

et al. 2017

Self Cite

View full text Add to dashboard Cite

Recent treatments of leukemias with chimeric antigen receptor (CAR) expressing T cells underline their impressive therapeutic potential. However, once adoptively transferred into patients, there is little scope left to shut them down after elimination of tumor cells or in case adverse side effects occur. This becomes of special relevance if they are directed against commonly expressed tumor associated antigens (TAAs) such as receptors of the ErbB family. To overcome this limitation, we recently established a modular CAR platform technology termed UniCAR. UniCARs are not directed against TAAs but instead against a unique peptide epitope on engineered recombinant targeting modules (TMs), which guide them to the target. In the absence of a TM UniCAR T cells are inactive. Thus an interruption of any UniCAR activity requires an elimination of unbound TM and the TM complexed with UniCAR T cells. Elimination of the latter one requires a disassembly of the UniCAR-TM complexes. Here, we describe a first nanobody (nb)-based TM directed against EGFR. The novel TM efficiently retargets UniCAR T cells to EGFR positive tumors and mediates highly efficient target-specific and target-dependent tumor cell lysis both in vitro and in vivo. After radiolabeling of the novel TM with 64 Cu and 68 Ga, we analyzed its biodistribution and clearance as well as the stability of the UniCAR-TM complexes. As expected unbound TM is rapidly eliminated while the elimination of the TM complexed with UniCAR T cells is delayed. Nonetheless, we show that UniCAR-TM complexes dissociate in vitro and in vivo in a concentration-dependent manner in line with the concept of a repeated stop and go retargeting of tumor cells via the UniCAR technology.

show abstract

Section: Expression and Purification Of Recombinant Antibodiesmentioning

confidence: 99%

“…[44][45][46][47] Recombinant Abs were detected by a mouse mAb directed to the his-tag (Qiagen) and a polyclonal rabbit a-mouse IgG conjugated with alkaline phosphatase (Dianova GmbH, Hamburg, Germany).…”

Section: Expression and Purification Of Recombinant Antibodiesmentioning

confidence: 99%

A novel nanobody-based target module for retargeting of T lymphocytes to EGFR-expressing cancer cells via the modular UniCAR platform

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…1). The potential of this modality has been demonstrated preclinically for both hematopoietic (2,3) and solid cancers (4)(5)(6)(7)(8). Tandem scFvs consisting of an anti-CD3 and an anti-TAA domain, termed bispecific T-cell engagers (BiTE), have advanced the furthest, with two agents under clinical investigation (9).…”

Section: Introductionmentioning

confidence: 99%

Redirected T-Cell Killing of Solid Cancers Targeted with an Anti-CD3/Trop-2–Bispecific Antibody Is Enhanced in Combination with Interferon-α

Rossi

Chang

et al. 2014

Molecular Cancer Therapeutics

View full text Add to dashboard Cite

Trop-2 has limited presence on normal tissues but is highly expressed in diverse epithelial cancers. (E1)-3s is a T-cell-redirecting trivalent bispecific antibody (bsAb), comprising an anti-CD3 scFv covalently linked to a stabilized dimer of a Trop-2-targeting Fab using Dock-and-Lock. We show for the first time that bsAbmediated bidirectional trogocytosis occurs between target and T cells and involves immunologic synapses. We studied the effects of interferon-a (INFa) on (E1)-3s-mediated T-cell killing of human gastric and pancreatic cancer cell lines. T-cell activation, cytokine induction, and cytotoxicity were evaluated ex vivo using peripheral blood mononuclear cells (PBMC) or T cells with NCI-N87 gastric cancer as target cells. In vivo activity was assayed with NCI-N87 and Capan-1 (pancreatic) xenografts. In the presence of target cells and PBMCs, (E1)-3s did not cause excess cytokine production. When combined with (E1)-3s, peginterferonalfa-2a-which alone did not increase T-cell activation or raise cytokine levels over baseline-increased CD69 expression but did not significantly increase cytokine induction. (E1) 3s mediated a highly potent T-cell lysis of NCI-N87 target cells in vitro. Inclusion of peginterferonalfa-2a or a more potent form of INFa, 20 Ã -2b, significantly potentiated the activity of (E1)-3s by more than 2.5-or 7-fold, respectively. In vivo, combining peginterferonalfa-2a with (E1)-3s delayed Capan-1 growth longer than each single agent. Similarly, combination therapy delayed tumor proliferation of NCI-N87 compared with (E1)-3s or peginterferonalfa-2a single-treatment groups. (E1)-3s effectively induced T-cell-mediated killing of Trop-2-expressing pancreatic and gastric cancers, which was enhanced with INFa.

show abstract

“…Subclasses of cytotoxic/cytolytic lymphocytes are able to recognize and attack transforming stem cells. In order to attack prostate stem cell antigenexpressing senescent prostate cancer cells, the T cell receptor could be redirected by bispecific antibodies (410,411 (412). The preventive value of dutasteride is in doubt: indeed it has utterly failed to prevent high grade disease.…”

Section: Ebvmentioning

confidence: 99%

Molecular biology of oncogenic inflammatory processes. I. Non-oncogenic and oncogenic pathogens, intrinsic inflammatory reactions without pathogens, and microRNA/DNA interactions (Review)

Sinkovics

2011

Int J Oncol

View full text Add to dashboard Cite

Abstract. In some inflammasomes tumor cells are generated. The internal environment of the inflammasome is conducive to the induction of malignant transformation. Epigenetic changes initiate this process. The subverted stromal connective tissue cells act to promote and sustain the process of malignant trans formation. In its early stages, the premalignant cells depend on paracrine circuitries for the reception of growth factors. The ligands are derived from the connective tissue, and the receptors are expressed on the recipient premalignant cells. The initial events are not a direct attack on the protooncogenes, and thus it may be entirely reversible. Epigenetic processes of hypermethylation of the genes at the promoters of tumor suppressor genes (to silence them), and deacetylation of the histones aimed at the promoters of protooncogenes (to activate them) are ongoing. A large number of short RNA sequences (interfering, micro, short hairpin, noncoding RNAs) silence tumor suppressor genes, by neutralizing their mRNAs. In a serial sequence oncogenes undergo amplifications, pointmutations, translocations and fusions. In its earliest stage, the process is reversible by demethylation of the silenced suppressor gene promoters (to reactivate them), or reacetylation of the histones of the oncogene promoters, thus deactivating them. The external administration of histone deacetylase inhibitors usually leads to the restoration of histone acetylation. In time, the uncorrected processes solidify into constitutive and irreversible gene mutations. Some of the pathogens inducing inflammations with consquential malignant transformation contain oncogenic gene sequences (papilloma viruses, EpsteinBarr virus, Kaposi's sarcomaassociated herpesvirus, hepatitis B and C viruses, Merkel cell polyoma virus, Helicobacter pylori, enterotoxigenic Bacteroides fragilis). These induced malignancies may be multifocal. Other pathogens are devoid of any known oncogenic genomic sequences (mycoplasma vavcarcinogenesis, chlamydia MALTlymphoma genesis). In these cases the host's inflammatory reactions induce the malignant transformation in serial sequences of gene alterations initiated by hypoxia and reactive oxygen and nitrogen species generation. Carcinogenic intrinsic inflammatory processes endogenously initiated without a pathogen are recognized. Chronic inflammatory processes signal the RNA/DNA complex. In response, the DNA may revert into its ancient primordial 'immortal' format, which the clinics recognize as 'oncogenesis'. The DNA remains the ultimate master of bioengineering in order to sustain life. A discussion on the most versatile and resistant primordial RNA/ DNA complex and the pre, proto, and unicellular world in which they coexisted is included. Contents1. Pathogens without oncogenic genomic sequences activating cellular oncogenes 2. The inflammasome 3. Tumor cell colonies generated in the inflammasomes 4. Pathogens with potentially oncogenic genomic sequences 5. Mechanisms of inflammatory carcinogenesis Pathogens without oncogenic...

show abstract

Retargeting of T cells to prostate stem cell antigen expressing tumor cells: Comparison of different antibody formats

Cited by 62 publications

References 29 publications

A novel nanobody-based target module for retargeting of T lymphocytes to EGFR-expressing cancer cells via the modular UniCAR platform

A novel nanobody-based target module for retargeting of T lymphocytes to EGFR-expressing cancer cells via the modular UniCAR platform

Redirected T-Cell Killing of Solid Cancers Targeted with an Anti-CD3/Trop-2–Bispecific Antibody Is Enhanced in Combination with Interferon-α

Molecular biology of oncogenic inflammatory processes. I. Non-oncogenic and oncogenic pathogens, intrinsic inflammatory reactions without pathogens, and microRNA/DNA interactions (Review)

Contact Info

Product

Resources

About