Table 2 Preclinical studies of cell therapies for RCC treatment

TCR gene-modified T cells

Design: T cells transduced with 5T4-directed scFv CD3ζ receptor.

Test: in vitro − 2220R, 2246R, 2245R cell lines.

T cells can be directed against the 5T4 protein, inducing cytotoxicity.

Griffiths et al. 2005 [22]

Design: High-avidity CD8 + T cell clones specific for an HLA-A2-restricted 5T4 epitope.

Test: in vitro - A498, BB65, LB1828 and DOBSKI cell lines.

TCR-engineered 5T4p17-specific CD8+ T cells can induce cytotoxicity of 5T4+ RCC cell lines.

Xu et al. 2019 [23]

Design: TCR53/PD-1:28 \(tm\)-transgenic T cells targeting PD-L1 and PD-L2.

Test: in vitro - RCC-26 and RCC-53 (high PD-L1 expression) cell lines.

PD-1:28 engineered T cells secreted significantly more IFN-γ, suggesting a beneficial combination with other therapeutic strategies.

Schlenker et al. 2017 [24]

CAR-T cells

Design: Two humanized CAIX-directed CAR constructs:

First generation Anti-CAIX G36 scFv-CD8-CD3ζ (CD8ζ) and

Second-generation Anti-CAIX G36 scFv-CD28-CD3ζ (CD28ζ).

Test: in vitro - skrc-52 (CAIX+), skrc-59 (CAIX-) cell lines

in vivo - nude mice inoculated with the cell lines.

Second-generation G36-CD28z CAR-T cells have in vivo antitumor responses .

Lo et al. 2014 [25]

Design: Two humanized CAIX-directed CAR constructs:

Anti-CAIX G36 scFv-CD28-CD3ζ (G36-CD28ζ) able to release anti-PD-L1 IgG1 or IgG4 using a bicistronic vector.

Test: in vitro - skrc-59 (CAIX+ PD-L1+) and skrc-59 (CAIX- PD-L1-) cell lines

in vivo - NSG mice inoculated with the cell lines.

The G36 Anti-CAIX CAR-T cells secreting human anti-PD-L1 antibodies in the ccRCC milieu boosted anti-tumor responses against RCC combating T cell exhaustion.

Suarez et al. 2016 [26]

Design: Two humanized CAIX-directed CAR construct:

Anti-CAIX G36 scFv-CD28-CD3ζ (G36-CD28ζ) and Anti-CAIX G36 scFv-41BB-CD3ζ (G36-41BBζ) able to release anti-PD-L1 IgG4 using a bicistronic vector.

Test: in vitro – skrc-59 (CAIX+ PD-L1+) and skrc-59 (CAIX- PD-L1-) cell lines

in vivo - NSG mice inoculated with the cell lines.

Anti-CAIX G36 CD28 CAR-T cells releasing anti-PD-L1 IgG4 antibodies offered exciting new prospects for the treatment of refractory ccRCC and hypoxic tumors.

Campos et al. 2022 [27]

Design: Comparison of second-generation humanized anti-CAIX G36- scFv CD28- CD3ζ and anti-CAIX G36-41BB CD3ζ) with a third-generation CAR (anti-CAIX G36-CD28-41BB- CD3ζ) using different CD4/CD8 proportions.

Test: in vitro - skrc-59 (CAIX+) cell line.

in vivo - NSG-SGM3 mice inoculated with the cell lines.

Anti-CAIX BBζ CAR4/8 CAR-T cells have the potential to be translated to clinic for treatment of ccRCC due to complete tumor regression.

Wang et al. 2021 [28]

Design: CAIX-directed CAR-T cells composed by a mouse anti-human CAIX-scFv, 4-1BB-CD3 ζ CAR in combination with the TKI sunitinib.

Test: in vitro - Ketr-3 and OSRC-2 cell lines.

in vivo - NOG mice inoculated with the cell lines.

Combination therapy with CAIX-CAR-T and sunitinib showed synergistic efficacy in a mouse lung metastasis model of human RCC.

Li et al. 2020 [29]

Design: PARPi olaparib (OLA) associated with CD70 directed-CAR-T 4-1BB CD3ζ.

Test: in vitro - 786-0, A498, and 769-P cell lines.

in vivo - NDG mice inoculated with 786-0 cell line.

This study indicates that the combination of CAR-T cell therapy with PARPi represents a potential therapeutic approach for RCC.

Ji et al. 2021 [30]

Design: Analysis of multiple anti-CD70 scFvs 4-1BB CD3ζ

allogeneic CAR-T cells associated with ritumixab.

Test: in vitro - 786-0 (CD70+++), ACHN (CD70++) and REH (CD70+) cell lines.

in vivo - NSG mice inoculated with the 786-0 cell line.

These efficacy data supported the evaluation of CD70 CAR-T cells for the treatment of RCC and has led to the advancement of an allogeneic CD70 CAR-T cell candidate into Phase I clinical trials.

Panowski et al. 2022 [31]

Design: c-met-directed third generation CAR-T cells containing CD28, 4-1BB and CD3ζ, in combination with axitinib.

Test: in vitro - A498 (c-met+++) and KMS11 cell line.

in vivo - NSG mice inoculated with A498-Luc cell line.

This study demonstrated the potential of anti-c-met CAR-T cell alone or in association with axitinib against RCC.

Mori et al. 2021 [32]

CAR-NK cells

Design: CAR based on anti HER2 scFv FRP5-CD28- CD3ζ (CAR 5.28.ζ ).

Test: in vitro - Murine Renca-lacZ/HER2, Renca-lacZ/EGFR cells lines.

in vivo - NSG implanted with Renca-lacZ/HER2 cell line.

NK-92/5.28ζ has antitumor activity resulting in fewer lung metastases compared to control.

Schonfeld et al. 2014 [33]

Design: Third-generation CAR‑NK92 cells (anti-CAIX scFv-CD28-41BB-CD3ζ) alone or combined with bortezomib.

Test: in vitro - OSRC-2, Ketr-3, ACHN and 293 cell lines.

in vivo - NSG mice implanted with Ketr-3luc+ cell line.

CAIX-Specific NK92 cells alone decreased RCC volume and the association with bortezomib boosted their antitumor effects, leading to complete remission in mice.

Zhang et al. 2018 [34]

Design: Third-generation CAR based in an EGFR-scFv-CD28-4-1BB-CD3ζ associated with cabozantinib.

Test: in vitro - 786-O, ACHN, Ketr-3 and OSRC-2 cell lines.

in vivo - ACHN-Luc implanted in NSG mice.

EGFR-directed CAR-NK92 cells decreased EGFR+ RCC, and the association with cabozantinib boosted the antitumor effects of the anti-EGFR CAR-NK92 cells.

Zhang et al. 2017 [35]

Genetic engineered NK cells

Design: NK cells transduced with human CXCR2.

Test: in vitro - ACHN, Caki-2, A498 cell lines.

CXCR2-transduced NK cells showed increased adhesion properties and ability to migrate along a CXCR2 ligands gradient.

Kremer et al. 2017 [36]

γδ T cells

Design: IL-15-induced γδ T cell compared with IL-2-γδ T cell.

Test: in vitro - 786-O, ACHN, Caki-1 cell lines.

in vivo - Patient-derived xenografts, NOG mice.

These results indicate that γδ T cells induced by IL-15 are more potent against RCC compared to IL-2-induced γδ T cells.

Zhang et al. 2021 [37]

Design: CD3 low Vγ9+-δ1+ TILs and peripheral blood Vγ9+-δ2 + T cells.

Test: in vitro - Caki-1 and ACHN cell lines.

Vγ9Vδ1 T cells induced cytotoxicity of RCC cells.

Lee et al. 2021 [38]

CAR-T cells

and oncolytic adenovírus

Design: Oncolytic adenovirus carrying decorin (OAV-DEC) with a mouse anti-human CAIX scFv-CAR-T 4-1BB-CD3ζ.

Test: in vitro - OSRC-2 (CAIX+++), 786-0 (CAIX++) and ACHN (CAIX+) cell lines.

in vivo - NSG mice using the OSRC-2 cell line.

Combined use of OAV-Decorin and CAIX targeted CAR-T displayed synergistic antitumor effects in vitro and in vivo by enhancing T cell persistence.

Zhang et al. 2022 [39]