Title: Chimeric antigen receptor-T cells targeting epithelial cell adhesion molecule antigens are
effective in the treatment of colorectal cancer
Version: 1 Date: 08 Jan 2024
Reviewer's report:
This  manuscript demonstrates new CAR T cells against EpCAM inhibiting the tumor growth
of colorectal tumor cells in vitro and in vivo. The experiments are well done and the manuscript
should be published.  Nevertheless some points have to be answered and discussed :
Results after abstract: please add peripheral blood mononuclear cells were extracted from
whome ? patients or mice ?
Conclusion: Last sentence: und can also better inhibit the growth and metastasis of colerectal
cancer in vitro and in vivo, better as what ?
In the introduction  you have to mention and to write about TCR engineered T cells, CAR T-
cells, TCR CARs and TCR like CARs which are all promising.  And please mention pro and
cons of these T cells. This is important for people that are not so familiar with the strategies
of T cell therapy. TCR engineered T cells, TCR CARs and TCR like CARs are able to target
intracellular neoantigens, which are specifically expressed only on tumor cells and not on normal
cells. Exist there an advantage of T cell therapy over therapies with antibodies, for example
bipecific antibodies targeting CD3 and  tumor associated antigens like EpCAM compared with
the strategy demonstrated in the manuscript.
Results:from where is the anti-scFv derived ? please add citation.
Can you please explain the amount of CD8, CD4 and CD3 positive cells found by FACs. What
is the amount of double positive CD4 and CD3 and double positive CD8 and CD3 cells? Could
you please  quantify the Ki67 experiments, Fig. 5F.
Discussion: Solid tumors are different from hematologic tumors, please explaine the differences
in more detail. Why do the T cell group injection of commun T cells have also an effect on tumor
growth and surveillance of mice ?
Conclusion: How can off/target effects by CAR T cells be prevented?  for example, Dual CARs
or Tandem CARs or synNotch CARs.
Please describe briefly the fourth generation CARs (Michael Hiltensperger and Angela M.
Krackhardt, Frontiers in Immunology 2023.
Title: Chimeric antigen receptor-T cells targeting epithelial cell adhesion molecule antigens are
effective in the treatment of colorectal cancer
Version: 1 Date: 08 Jan 2024
Reviewer's report:
This  manuscript demonstrates new CAR T cells against EpCAM inhibiting the tumor growth
of colorectal tumor cells in vitro and in vivo. The experiments are well done and the manuscript
should be published.  Nevertheless some points have to be answered and discussed :
Results after abstract: please add peripheral blood mononuclear cells were extracted from
whome ? patients or mice ?
Conclusion: Last sentence: und can also better inhibit the growth and metastasis of colerectal
cancer in vitro and in vivo, better as what ?
In the introduction  you have to mention and to write about TCR engineered T cells, CAR T-
cells, TCR CARs and TCR like CARs which are all promising.  And please mention pro and
cons of these T cells. This is important for people that are not so familiar with the strategies
of T cell therapy. TCR engineered T cells, TCR CARs and TCR like CARs are able to target
intracellular neoantigens, which are specifically expressed only on tumor cells and not on normal
cells. Exist there an advantage of T cell therapy over therapies with antibodies, for example
bipecific antibodies targeting CD3 and  tumor associated antigens like EpCAM compared with
the strategy demonstrated in the manuscript.
Results:from where is the anti-scFv derived ? please add citation.
Can you please explain the amount of CD8, CD4 and CD3 positive cells found by FACs. What
is the amount of double positive CD4 and CD3 and double positive CD8 and CD3 cells? Could
you please  quantify the Ki67 experiments, Fig. 5F.
Discussion: Solid tumors are different from hematologic tumors, please explaine the differences
in more detail. Why do the T cell group injection of commun T cells have also an effect on tumor
growth and surveillance of mice ?
Conclusion: How can off/target effects by CAR T cells be prevented?  for example, Dual CARs
or Tandem CARs or synNotch CARs.
Please describe briefly the fourth generation CARs (Michael Hiltensperger and Angela M.
Krackhardt, Frontiers in Immunology 2023.
Please review this paper.
user
Title: Chimeric antigen receptor-T cells targeting epithelial cell adhesion molecule antigens are
effective in the treatment of colorectal cancer
Version: 1 Date: 08 Jan 2024
Reviewer's report:
This  manuscript demonstrates new CAR T cells against EpCAM inhibiting the tumor growth
of colorectal tumor cells in vitro and in vivo. The experiments are well done and the manuscript
should be published.  Nevertheless some points have to be answered and discussed :
Results after abstract: please add peripheral blood mononuclear cells were extracted from
whome ? patients or mice ?
Conclusion: Last sentence: und can also better inhibit the growth and metastasis of colerectal
cancer in vitro and in vivo, better as what ?
In the introduction  you have to mention and to write about TCR engineered T cells, CAR T-
cells, TCR CARs and TCR like CARs which are all promising.  And please mention pro and
cons of these T cells. This is important for people that are not so familiar with the strategies
of T cell therapy. TCR engineered T cells, TCR CARs and TCR like CARs are able to target
intracellular neoantigens, which are specifically expressed only on tumor cells and not on normal
cells. Exist there an advantage of T cell therapy over therapies with antibodies, for example
bipecific antibodies targeting CD3 and  tumor associated antigens like EpCAM compared with
the strategy demonstrated in the manuscript.
Results:from where is the anti-scFv derived ? please add citation.
Can you please explain the amount of CD8, CD4 and CD3 positive cells found by FACs. What
is the amount of double positive CD4 and CD3 and double positive CD8 and CD3 cells? Could
you please  quantify the Ki67 experiments, Fig. 5F.
Discussion: Solid tumors are different from hematologic tumors, please explaine the differences
in more detail. Why do the T cell group injection of commun T cells have also an effect on tumor
growth and surveillance of mice ?
Conclusion: How can off/target effects by CAR T cells be prevented?  for example, Dual CARs
or Tandem CARs or synNotch CARs.
Please describe briefly the fourth generation CARs (Michael Hiltensperger and Angela M.
Krackhardt, Frontiers in Immunology 2023.
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