Center for Molecular Medicine Cologne

Hinrich Abken - assoc. RG

Metabolic addiction and reprogramming of an anti-tumor T cell response

Summary

The spectacular success of chimeric antigen receptor (CAR) T cells in treating hematologic malignancies opens new ways in cancer therapy; the impact of metabolic reprogramming within the tumor tissue on the T cell anti-tumor response remains unclear. We address how metabolic reprogramming alters T cell physiology, persistence and anti-tumor response and whether this can be used to improve the T cell response towards an enduring control of malignant diseases.

Introduction 

Adoptive cell therapy with the transfer of genetically redirected autologous T cells has shown spectacular success in the treatment of hematologic malignancies and some solid tumors. Genetic engineering with a chimeric antigen receptor (CAR) is commonly used for redirecting T cells towards defined target. As a recombinant, composite membrane receptor the CAR recognizes a surface antigen on the targeted cancer cell by an antibody-derived extracellular domain and activates by a T cell receptor (TCR) derived intracellular signalling domain. A major advantage of the CAR T cell therapy is the applicability to a wide range of patients, based on the antibody mediated, MHC-independent recognition of antigen, to a wide range of malignancies due to targeting various tumor-associated antigens, and the orchestrated T cell activation based on signalling through the combination of signalling moieties. Of note, the modular CAR design recapitulates many aspects of the natural T cell signalling machinery and redirects the T cell response in a defined fashion.

Antigen receptor-redirected T cells derived from hematopoietic precursor cells lack expression of the endogenous TCR/CD3 receptor and exhibit specific anti-tumor capacities.

Currently, adoptive T cell therapy is derived from autologous peripheral blood T cells; T cells derived from healthy allogeneic donors may have advantages, however, the expected occurrence of graft versus host disease as a consequence of the diverse allogeneic T cell receptor (TCR) repertoire expressed by these cells seriously compromises the approach. We generated T cells from cord blood hematopoietic progenitor cells that were transduced to express either a CAR or a TCR with or without built-in costimulatory sequence. Transgenic precursor cells were in vitro differentiated to CD5+ CD7+ positive T lineage precursors, to CD4+ CD8+ double positive cells and finally to mature AR+ T cells on an OP9-DL1 feeder layer. The receptor expressing T cells were largely of naive CD45RA+CD62L+ phenotype and lack CD3 indicating that rearrangements of the endogenous TCR locus were blocked. However, the cells were functional as they displayed specific cytotoxic activity and cytokine release. Data sustain the concept that cord blood derived, in vitro generated CD3- CAR T cells can be used to more effectively eliminate tumor leukemic cells while at the same time limiting the occurrence of graft versus host disease.
Van Caeneghem et al., OncoImmunol. (2017)

CD4+CD25- T cells, but not CD4+CD25+ Treg cells, are cytolytic when redirected by a CAR.

Evidences are accumulating that CD4+ T cells can physiologically mediate antigen specific target cell lysis. By circumventing MHC restriction through an engineered CAR CD4+ T cells lyse defined target cells as efficiently as do CD8+ T cells. For activation, Treg cells require a strong CD28 signal together with CD3ζ inducing a distinct cytokine pattern with high IL-10 and lack of IL-2 release. Despite strong and antigen-specific activation, CAR Treg cells produced only weak target cell lysis whereas CD4+CD25- CAR T cells were potent killers. Cytolysis did not correlate with the target cell sensitivity to Fas/FasL mediated killing; CD4+CD25- T cells upregulated perforin and granzyme B upon CAR activation whereas Treg cells did less. The analysis clearly indicates different cytolytic capacities of CAR redirected conventional CD4+ cells and Treg cells which warrants further evaluation.
Hombach et al., Cancers (2017)

Chimeric antigen receptor-redirected regulatory T cells suppress experimental allergic airway inflammation, a model of asthma.

We explored a regulatory T (Treg) cell-based therapy in the treatment of allergic inflammation using a model for asthma characterized by a chronic, Th2 cell dominated immune response to allergen. We redirected Tregs by a CAR towards lung epithelia in mice upon experimentally induced allergic asthma, closely mimicking the clinical situation. Adoptively transferred CAR Tregs accumulated in the lung and in regional lymph nodes, reduced hyper-reactivity and diminished inflammation, prevented excessive pulmonary mucus production and increase in IgE and Th2 cytokine serum levels. CAR Tregs were more efficient in controlling asthma than non-modified Tregs, paving the way for CAR Treg cell therapy of severe allergic asthma.
Skuljec et al., Frontiers Immunol. (2017)

IL-18 induces T-bet(high) FoxO1(low) T cells with a distinct signature for the CAR therapy of advanced solid tumors.

We explored to sustain acute inflammation while preventing exhaustion by modulating the metabolic and signaling signature of effector T cells. A systematic screen identified IL-18 as a pro-inflammatory response modifier converting CAR T cells towards T-bet(high) FoxO1(low) effector cells resisting exhaustion. CAR T cells with inducible IL-18 release showed a distinct cytokine and functional signature superior in the activity against large established tumors. IL-18 CAR T cell treatment was accompanied by an overall change in the tumor immune cell landscape. 

T cells engineered with a TCR and transgenic inducible IL-12 and IL-18, respectively, produced enhanced levels of IFN-γ. Adoptive transfer of T cells with a melanoma specific TCR and inducible IL-12 to melanoma-bearing mice resulted in severe, edema-like toxicity that was accompanied by enhanced inflammation and infiltration of macrophages into the tumor. In contrast, transfer of  IL-18 producing TCR T cells were safe and significantly reduced tumor burden, prolonged overall survival. Data imply “IL18 TRUCKs” for eradicating large solid tumor lesions in an advanced stage of the disease.
Chmielewski et al., Cell Reports (2017)
Kunert et al., OncoImmunol (2018)

T cells with CD28-ζ CAR resist TGF-β repression through IL-2 signaling.

We revealed that T cells with a CD28 CAR, but not with a 4-1BB CAR, resist TGF-β repression due to LCK activation and autocrine IL-2 receptor signaling. Deleting the LCK binding motif in the CD28 CAR abolished both IL-2 secretion and TGF-β resistance. Other γ-cytokines like IL-7 and IL-15 could replace IL-2 in this context resulting in resistant T cells engineered with IL-2 deficient CD28 CAR and a hybrid IL-7 receptor to provide IL-2R β-chain signaling upon IL-7 binding. Data draw the concept that an autocrine loop resulting in IL-2R signaling can make CAR T cells more potent in staying active against TGF-β+ solid tumors.
Golumba-Nagy et al., Mol. Ther. (2018)

Aleksandrova, K., Leise, J., Priesner, C., Melk, A., Kubaink, F., Abken, H., Hombach, A., Aktas, M., Essl, M., Burger, I., Kaiser, A., Rauser, G., Jurk, M., Goudeva, L., Glienke, W., Arseniev, L., Esser, R., and Kohl, U. (2019). Functionality and Cell Senescence of CD4/ CD8-Selected CD20 CAR T Cells Manufactured Using the Automated CliniMACS Prodigy(R) Platform. Transfus Med Hemother 46, 47-54.

Chmielewski, M., Kuehle, J., Chrobok, D., Riet, N., Hallek, M., and Abken, H. (2019). FimH-based display of functional eukaryotic proteins on bacteria surfaces. Sci Rep 9, 8410.

Hombach, A.A., Rappl, G., and Abken, H. (2019). Blocking CD30 on T Cells by a Dual Specific CAR for CD30 and Colon Cancer Antigens Improves the CAR T Cell Response against CD30(-) Tumors. Mol Ther10.1016/j.ymthe.2019.06.007.

 

Bergmeier V, Etich J, Pitzler L, Frie C, Koch M, Fischer M, Rappl G, Abken H, Tomasek JJ, and Brachvogel B (2018). Identification of a myofibroblast-specific expression signature in skin wounds. Matrix Biol 65, 59-74.

Bluhm J, Kieback E, Marino SF, Oden F, Westermann J, Chmielewski M, Abken H, Uckert W, Hopken UE, and Rehm A (2018). CAR T Cells with Enhanced Sensitivity to B Cell Maturation Antigen for the Targeting of B Cell Non-Hodgkin's Lymphoma and Multiple Myeloma. Mol Ther 26, 1906-1920.

De Munter S, Ingels J, Goetgeluk G, Bonte S, Pille M, Weening K, Kerre T, Abken H, and Vandekerckhove B (2018). Nanobody Based Dual Specific CARs. Int J Mol Sci 19.

Golumba-Nagy V, Kuehle J, Hombach AA, and Abken H (2018). CD28-zeta CAR T Cells Resist TGF-beta Repression through IL-2 Signaling, Which Can Be Mimicked by an Engineered IL-7 Autocrine Loop. Mol Ther 26, 2218-2230.

Kunert A, Chmielewski M, Wijers R, Berrevoets C, Abken H, and Debets R (2018). Intra-tumoral production of IL18, but not IL12, by TCR-engineered T cells is non-toxic and counteracts immune evasion of solid tumors. Oncoimmunology 7.

Probst K, Stermann J, von Bomhard I, Etich J, Pitzler L, Niehoff A, Bluhm B, Xu HC, Lang PA, Chmielewski M, Abken H, Blissenbach B, Machova A, Papadopoulou N, and Brachvogel B (2018). Depletion of Collagen IX Alpha1 Impairs Myeloid Cell Function. Stem Cells10.1002/stem.2892.

Kohl U, Arsenieva S, Holzinger A, and Abken H (2018). CAR T Cells in Trials: Recent Achievements and Challenges that Remain in the Production of Modified T Cells for Clinical Applications. Hum Gene Ther 29, 559-568.

Abken H (2017). Driving CARs on the Highway to Solid Cancer: Some Considerations on the Adoptive Therapy with CAR T Cells. Hum Gene Ther 28, 1047-60.

Bergmeier V, Etich J, Pitzler L, Frie C, Koch M, Fischer M, Rappl G, Abken H, Tomasek JJ, and Brachvogel B (2017). Identification of a myofibroblast-specific expression signature in skin wounds. Matrix Biol10.1016/j.matbio.2017.07.005.

Chmielewski M, and Abken H (2017). CAR T Cells Releasing IL-18 Convert to T-Bet(high) FoxO1(low) Effectors that Exhibit Augmented Activity against Advanced Solid Tumors. Cell Rep 21, 3205-19.

Golumba-Nagy V, Kuehle J, and Abken H (2017). Genetic Modification of T Cells with Chimeric Antigen Receptors: A Laboratory Manual. Hum Gene Ther Methods10.1089/hgtb.2017.083.

Herling M, Rengstl B, Scholtysik R, Hartmann S, Kuppers R, Hansmann ML, Diebner HH, Roeder I, Abken H, Newrzela S, and Kirberg J (2017). Concepts in mature T-cell lymphomas - highlights from an international joint symposium on T-cell immunology and oncology. Leuk Lymphoma 58, 788-96.

Holzinger A, and Abken H (2017). CAR T cells targeting solid tumors: carcinoembryonic antigen (CEA) proves to be a safe target. Cancer Immunol Immunother 66, 1505-7.

Hombach AA, and Abken H (2017a). Most Do, but Some Do Not: CD4(+)CD25(-) T Cells, but Not CD4(+)CD25(+) Treg Cells, Are Cytolytic When Redirected by a Chimeric Antigen Receptor (CAR). Cancers (Basel) 9.

Hombach AA, and Abken H (2017b). Shared target antigens on cancer cells and tissue stem cells: go or no-go for CAR T cells? Expert Rev Clin Immunol 13, 151-5.

Martyniszyn A, Krahl AC, Andre MC, Hombach AA, and Abken H (2017). CD20-CD19 bispecific CAR T cells for the treatment of B cell malignancies. Hum Gene Ther10.1089/hum.2017.126.

Sabour D, Srinivasan SP, Rohani S, Wagh V, Gaspar JA, Panek D, Ardestani MA, Doss MX, Riet N, Abken H, Hescheler J, Papadopoulos S, and Sachinidis A (2017). STRIP2 Is Indispensable for the Onset of Embryonic Stem Cell Differentiation. Mol Ther Methods Clin Dev 5, 116-29.

Shimabukuro-Vornhagen A, Garcia-Marquez M, Fischer RN, Iltgen-Breburda J, Fiedler A, Wennhold K, Rappl G, Abken H, Lehmann C, Herling M, Wolf D, Fatkenheuer G, Rubbert-Roth A, Hallek M, Theurich S, and von Bergwelt-Baildon M (2017). Antigen-presenting human B cells are expanded in inflammatory conditions. J Leukoc Biol 101, 577-87.

Skuljec J, Chmielewski M, Happle C, Habener A, Busse M, Abken H, and Hansen G (2017). Chimeric Antigen Receptor-Redirected Regulatory T Cells Suppress Experimental Allergic Airway Inflammation, a Model of Asthma. Front Immunol 8, 1125.

Van Caeneghem Y, De Munter S, Tieppo P, Goetgeluk G, Weening K, Verstichel G, Bonte S, Taghon T, Leclercq G, Kerre T, Debets R, Vermijlen D, Abken H, and Vandekerckhove B (2017). Antigen receptor-redirected T cells derived from hematopoietic precursor cells lack expression of the endogenous TCR/CD3 receptor and exhibit specific antitumor capacities. Oncoimmunology 6, e1283460.

Wennhold K, Thelen M, Schlosser HA, Haustein N, Reuter S, Garcia-Marquez M, Lechner A, Kobold S, Rataj F, Utermohlen O, Chakupurakal G, Theurich S, Hallek M, Abken H, Shimabukuro-Vornhagen A, and von Bergwelt-Baildon M (2017). Using Antigen-Specific B Cells to Combine Antibody and T Cell-Based Cancer Immunotherapy. Cancer Immunol Res 5, 730-43.

 

Prof. Dr. Hinrich Abken CMMC Cologne
Prof. Dr. Hinrich Abken

CMMC Affiliation

associated RG (05/2018-06/2019) Principal Investigator B 01 (1/2017-05/2018)

+49 941 943 5980

+49 941 943 5932

CMMC Affiliation

present address: Am Biopark 9

93053 Regensburg

http://www.rcii.de/forschung/lehrstuehle/lehrstuhl-fuer-gen-immuntherapie/

CMMC Profile Page

Curriculum Vitae (CV)

Publications - Hinrich Abken

Link to PubMed

Group Members

Post Docs:

  • Markus Chmielewski
  • Andreas Hombach
  • Johannes Kühle

Technicians:

  • Danuta Chrobok
  • Nicole Hoffmann
  • Petra Hofmann
  • Birgit Hops