Cytotoxic effect of CLL‑1 CAR‑T cell immunotherapy with PD‑1 silencing on relapsed/refractory acute myeloid leukemia

Lin,Guoqiang; Zhang,Yanming; Yu,Lei; Wu,Depei

doi:10.3892/mmr.2021.11847

Cytotoxic effect of CLL‑1 CAR‑T cell immunotherapy with PD‑1 silencing on relapsed/refractory acute myeloid leukemia

Authors:
- Guoqiang Lin
- Yanming Zhang
- Lei Yu
- Depei Wu
View Affiliations

Affiliations: National Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China, Department of Hematology, Huai'an Hospital Affiliated to Xuzhou Medical College, Huai'an Second People's Hospital, Huai'an, Jiangsu 223002, P.R. China, Institute of Biomedical Engineering and Technology, Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200065, P.R. China
Published online on: January 18, 2021 https://doi.org/10.3892/mmr.2021.11847
Article Number: 208
Copyright: © Lin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The activation of chimeric antigen receptor (CAR)‑T cells can lead to persistently high levels of programmed cell death 1 (PD‑1) antigen and eventually causes the exhaustion of T cells. The effectiveness of CAR‑T cells targeting C‑type lectin‑like molecule‑1 (CLL‑1) combined with PD‑1 silencing therapy for acute myeloid leukemia (AML) was evaluated in the present study. CLL‑1 levels in primary AML bone marrow samples was examined using flow cytometric analysis. We designed a CLL‑1 CAR‑T, containing CLL‑1‑specific single‑chain variable fragment, CD28, OX40, CD8 hinge and TM and CD3‑ζ signaling domains. CLL‑1 CAR‑T with PD‑1 silencing was constructed. It was confirmed that CLL‑1 is expressed on the surface of AML cells. CLL‑1 CAR‑T showed specific lysing activity against CLL‑1⁺ AML cells. PD‑1 silencing enhanced the killing ability of CLL‑1 CAR‑T. Furthermore, it was found that CAR‑T derived from healthy donor T cells was more effective in killing THP‑1 cells (a human acute monocytic leukemia cell line) than those from patient‑derived T cells. These results indicated that CLL‑1 CAR‑T and PD‑1 knockdown CLL‑1 CAR‑T could be used as a potential immunotherapy to treat relapsed or refractory AML.

View Figures

View References

1	Juliusson G, Lazarevic V, Hörstedt AS, Hagberg O and Höglund M; Swedish Acute Leukemia Registry Group, : Acute myeloid leukemia in the real world: Why population-based registries are needed. Blood. 119:3890–3899. 2012. View Article : Google Scholar : PubMed/NCBI
2	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
3	Shallis RM, Wang R, Davidoff A, Ma X and Zeidan AM: Epidemiology of acute myeloid leukemia: Recent progress and enduring challenges. Blood Rev. 36:70–87. 2019. View Article : Google Scholar : PubMed/NCBI
4	National Cancer Institute: Surveillance, Epidemiology, and End Results (SEER) Program Cancer Stat Facts: Leykemia-Acute myeloid leukemia (AML). https://seer.cancer.gov/statfacts/html/amyl.htmlApril 18–2019
5	Chen C, Wang P and Wang C: Prognostic nomogram for adult patients with acute myeloid leukemia: A SEER database analysis. Medicine (Baltimore). 98:e158042019. View Article : Google Scholar : PubMed/NCBI
6	Chen W, Zheng R, Zhang S, Zeng H, Xia C, Zuo T, Yang Z, Zou X and He J: Cancer incidence and mortality in China, 2013. Cancer Lett. 401:63–71. 2017. View Article : Google Scholar : PubMed/NCBI
7	Rodriguez-Abreu D, Bordoni A and Zucca E: Epidemiology of hematological malignancies. Ann Oncol. 18 (Suppl 1):i3–i8. 2007. View Article : Google Scholar : PubMed/NCBI
8	Sant M, Allemani C, Tereanu C, De Angelis R, Capocaccia R, Visser O, Marcos-Gragera R, Maynadié M, Simonetti A, Lutz JM, et al HAEMACARE Working Group, : Incidence of hematologic malignancies in Europe by morphologic subtype: Results of the HAEMACARE project. Blood. 116:3724–3734. 2010. View Article : Google Scholar : PubMed/NCBI
9	Wang X, Xiao Q, Wang Z and Feng WL: CAR-T therapy for leukemia: Progress and challenges. Transl Res. 182:135–144. 2017. View Article : Google Scholar : PubMed/NCBI
10	Tasian SK, Kenderian SS, Shen F, Ruella M, Shestova O, Kozlowski M, Li Y, Schrank-Hacker A, Morrissette JJD, Carroll M, et al: Optimized depletion of chimeric antigen receptor T-cells in murine xenograft models of human acute myeloid leukemia. Blood. 129:2395–2407. 2017. View Article : Google Scholar : PubMed/NCBI
11	Clarke CA and Glaser SL: Acute myeloid leukemia. N Engl J Med. 342:358–359. 2000. View Article : Google Scholar : PubMed/NCBI
12	Rowe JM and Tallman MS: How I treat acute myeloid leukemia. Blood. 116:3147–3156. 2010. View Article : Google Scholar : PubMed/NCBI
13	Stahl M, Kim TK and Zeidan AM: Update on acute myeloid leukemia stem cells: New discoveries and therapeutic opportunities. World J Stem Cells. 8:316–331. 2016. View Article : Google Scholar : PubMed/NCBI
14	Lichtenegger FS, Krupka C, Haubner S, Köhnke T and Subklewe M: Recent developments in immunotherapy of acute myeloid leukemia. J Hematol Oncol. 10:1422017. View Article : Google Scholar : PubMed/NCBI
15	Taussig DC, Pearce DJ, Simpson C, Rohatiner AZ, Lister TA, Kelly G, Luongo JL, Danet-Desnoyers GA and Bonnet D: Hematopoietic stem cells express multiple myeloid markers: Implications for the origin and targeted therapy of acute myeloid leukemia. Blood. 106:4086–4092. 2005. View Article : Google Scholar : PubMed/NCBI
16	Tettamanti S, Marin V, Pizzitola I, Magnani CF, Giordano Attianese GM, Cribioli E, Maltese F, Galimberti S, Lopez AF, Biondi A, et al: Targeting of acute myeloid leukaemia by cytokine-induced killer cells redirected with a novel CD123-specific chimeric antigen receptor. Br J Haematol. 161:389–401. 2013. View Article : Google Scholar : PubMed/NCBI
17	Ghaffari S, Smadja-Joffe F, Oostendorp R, Lévesque JP, Dougherty G, Eaves A and Eaves C: CD44 isoforms in normal and leukemic hematopoiesis. Exp Hematol. 27:978–993. 1999. View Article : Google Scholar : PubMed/NCBI
18	Kikushige Y, Shima T, Takayanagi S, Urata S, Miyamoto T, Iwasaki H, Takenaka K, Teshima T, Tanaka T, Inagaki Y, et al: TIM-3 is a promising target to selectively kill acute myeloid leukemia stem cells. Cell Stem Cell. 7:708–717. 2010. View Article : Google Scholar : PubMed/NCBI
19	Hosen N, Park CY, Tatsumi N, Oji Y, Sugiyama H, Gramatzki M, Krensky AM and Weissman IL: CD96 is a leukemic stem cell-specific marker in human acute myeloid leukemia. Proc Natl Acad Sci USA. 104:11008–11013. 2007. View Article : Google Scholar : PubMed/NCBI
20	Bonardi F, Fusetti F, Deelen P, van Gosliga D, Vellenga E and Schuringa JJ: A proteomics and transcriptomics approach to identify leukemic stem cell (LSC) markers. Mol Cell Proteomics. 12:626–637. 2013. View Article : Google Scholar : PubMed/NCBI
21	Saito Y, Kitamura H, Hijikata A, Tomizawa-Murasawa M, Tanaka S, Takagi S, Uchida N, Suzuki N, Sone A, Najima Y, et al: Identification of therapeutic targets for quiescent, chemotherapy-resistant human leukemia stem cells. Sci Transl Med. 2:17ra92010. View Article : Google Scholar : PubMed/NCBI
22	Moshaver B, van Rhenen A, Kelder A, van der Pol M, Terwijn M, Bachas C, Westra AH, Ossenkoppele GJ, Zweegman S and Schuurhuis GJ: Identification of a small subpopulation of candidate leukemia-initiating cells in the side population of patients with acute myeloid leukemia. Stem Cells. 26:3059–3067. 2008. View Article : Google Scholar : PubMed/NCBI
23	Liu J, Wang L, Zhao F, Tseng S, Narayanan C, Shura L, Willingham S, Howard M, Prohaska S, Volkmer J, et al: Pre-clinical development of a humanized anti-CD47 antibody with anti-cancer therapeutic potential. PLoS One. 10:e01373452015. View Article : Google Scholar : PubMed/NCBI
24	Amadori S, Suciu S, Selleslag D, Aversa F, Gaidano G, Musso M, Annino L, Venditti A, Voso MT, Mazzone C, et al: Gemtuzumab ozogamicin versus best supportive care in older patients with newly diagnosed acute myeloid leukemia unsuitable for intensive chemotherapy: Results of the randomized phase III EORTC-GIMEMA AML-19 trial. J Clin Oncol. 34:972–979. 2016. View Article : Google Scholar : PubMed/NCBI
25	Park JH, Rivière I, Gonen M, Wang X, Sénéchal B, Curran KJ, Sauter C, Wang Y, Santomasso B, Mead E, et al: Long-term follow-up of CD19 CAR therapy in acute lymphoblastic leukemia. N Engl J Med. 378:449–459. 2018. View Article : Google Scholar : PubMed/NCBI
26	Cao J, Wang G, Cheng H, Wei C, Qi K, Sang W, Zhenyu L, Shi M, Li H, Qiao J, et al: Potent anti-leukemia activities of humanized CD19-targeted chimeric antigen receptor T (CAR-T) cells in patients with relapsed/refractory acute lymphoblastic leukemia. Am J Hematol. 93:851–858. 2018. View Article : Google Scholar : PubMed/NCBI
27	Sotillo E, Barrett DM, Black KL, Bagashev A, Oldridge D, Wu G, Sussman R, Lanauze C, Ruella M, Gazzara MR, et al: Convergence of acquired mutations and alternative splicing of CD19 enables resistance to CART-19 immunotherapy. Cancer Discov. 5:1282–1295. 2015. View Article : Google Scholar : PubMed/NCBI
28	Fischer J, Paret C, El Malki K, Alt F, Wingerter A, Neu MA, Kron B, Russo A, Lehmann N, Roth L, et al: CD19 isoforms enabling resistance to CART-19 immunotherapy are expressed in B-ALL patients at initial diagnosis. J Immunother. 40:187–195. 2017. View Article : Google Scholar : PubMed/NCBI
29	Fan M, Li M, Gao L, Geng S, Wang J, Wang Y, Yan Z and Yu L: Chimeric antigen receptors for adoptive T cell therapy in acute myeloid leukemia. J Hematol Oncol. 10:1512017. View Article : Google Scholar : PubMed/NCBI
30	Beavis PA, Sek K and Darcy PK: A novel target antigen for the treatment of acute myeloid leukemia by CAR-T cells. Mol Ther. 25:1997–1998. 2017. View Article : Google Scholar : PubMed/NCBI
31	Prommersberger S, Jetani H, Danhof S, Monjezi R, Nerreter T, Beckmann J, Einsele H and Hudecek M: Novel targets and technologies for CAR-T cells in multiple myeloma and acute myeloid leukemia. Curr Res Transl Med. 66:37–38. 2018. View Article : Google Scholar : PubMed/NCBI
32	Lu H, Zhou Q, Deshmukh V, Phull H, Ma J, Tardif V, Naik RR, Bouvard C, Zhang Y, Choi S, et al: Targeting human C-type lectin-like molecule-1 (CLL1) with a bispecific antibody for immunotherapy of acute myeloid leukemia. Angew Chem Int Ed Engl. 53:9841–9845. 2014. View Article : Google Scholar : PubMed/NCBI
33	Bakker AB, van den Oudenrijn S, Bakker AQ, Feller N, van Meijer M, Bia JA, Jongeneelen MA, Visser TJ, Bijl N, Geuijen CA, et al: C-type lectin-like molecule-1: A novel myeloid cell surface marker associated with acute myeloid leukemia. Cancer Res. 64:8443–8450. 2004. View Article : Google Scholar : PubMed/NCBI
34	Wang J, Chen S, Xiao W, Li W, Wang L, Yang S, Wang W, Xu L, Liao S, Liu W, et al: CAR-T cells targeting CLL-1 as an approach to treat acute myeloid leukemia. J Hematol Oncol. 11:72018. View Article : Google Scholar : PubMed/NCBI
35	Tashiro H, Sauer T, Shum T, Parikh K, Mamonkin M, Omer B, Rouce RH, Lulla P, Rooney CM, Gottschalk S, et al: Treatment of acute myeloid leukemia with T-cells expressing chimeric antigen receptors directed to C-type lectin-like molecule-1. Mol Ther. 25:2202–2213. 2017. View Article : Google Scholar : PubMed/NCBI
36	Heczey A, Louis CU, Savoldo B, Dakhova O, Durett A, Grilley B, Liu H, Wu MF, Mei Z, Gee A, et al: CAR-T cells administered in combination with lymphodepletion and PD-1 inhibition to patients with neuroblastoma. Mol Ther. 25:2214–2224. 2017. View Article : Google Scholar : PubMed/NCBI
37	Ankri C, Shamalov K, Horovitz-Fried M, Mauer S and Cohen CJ: Human T-cells engineered to express a programmed death 1/28 costimulatory retargeting molecule display enhanced antitumor activity. J Immunol. 191:4121–4129. 2013. View Article : Google Scholar : PubMed/NCBI
38	Arber DA, Orazi A, Hasserjian R, Thiele J, Borowitz MJ, Le Beau MM, Bloomfield CD, Cazzola M and Vardiman JW: The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood. 127:2391–2405. 2016. View Article : Google Scholar : PubMed/NCBI
39	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
40	Jin HT, Ahmed R and Okazaki T: Role of PD-1 in regulating T cell immunity. Curr Top Microbiol Immunol. 350:17–37. 2011.PubMed/NCBI
41	van Rhenen A, van Dongen GA, Kelder A, Rombouts EJ, Feller N, Moshaver B, Stigter-van Walsum M, Zweegman S, Ossenkoppele GJ and Jan Schuurhuis G: The novel AML stem cell associated antigen CLL-1 aids in discrimination between normal and leukemic stem cells. Blood. 110:2659–2666. 2007. View Article : Google Scholar : PubMed/NCBI
42	Larsen HO, Roug AS, Just T, Brown GD and Hokland P: Expression of the hMICL in acute myeloid leukemia-a highly reliable disease marker at diagnosis and during follow-up. Cytometry B Clin Cytom. 82:3–8. 2012. View Article : Google Scholar : PubMed/NCBI
43	Darwish NH, Sudha T, Godugu K, Elbaz O, Abdelghaffar HA, Hassan EE and Mousa SA: Acute myeloid leukemia stem cell markers in prognosis and targeted therapy: Potential impact of BMI-1, TIM-3 and CLL-1. Oncotarget. 7:57811–57820. 2016. View Article : Google Scholar : PubMed/NCBI
44	Wang YY, Chen WL, Weng XQ, Sheng Y, Wu J, Hao J, Liu ZY, Zhu YM, Chen B, Xiong SM, et al: Low CLL-1 expression is a novel adverse predictor in 123 patients with de novo CD34⁺ acute myeloid leukemia. Stem Cells Dev. 26:1460–1467. 2017. View Article : Google Scholar : PubMed/NCBI
45	Laborda E, Mazagova M, Shao S, Wang X, Quirino H, Woods AK, Hampton EN, Rodgers DT, Kim CH, Schultz PG, et al: Development of a chimeric antigen receptor targeting C-type lectin-like molecule-1 for human acute myeloid leukemia. Int J Mol Sci. 18:182017. View Article : Google Scholar
46	Dunn GP, Old LJ and Schreiber RD: The immunobiology of cancer immunosurveillance and immunoediting. Immunity. 21:137–148. 2004. View Article : Google Scholar : PubMed/NCBI
47	Park Y, Lim J, Kim S, Song I, Kwon K, Koo S and Kim J: The prognostic impact of lymphocyte subsets in newly diagnosed acute myeloid leukemia. Blood Res. 53:198–204. 2018. View Article : Google Scholar : PubMed/NCBI
48	Alcasid M, Ma L, Gotlib JR, Arber DA and Ohgami RS: The clinicopathologic significance of lymphocyte subsets in acute myeloid leukemia. Int J Lab Hematol. 39:129–136. 2017. View Article : Google Scholar : PubMed/NCBI
49	Gray KD, Vedvyas Y, Kalloo O, Shevlin E and Min IM: Abstract 2738: PD-L1/PD-1 checkpoint inhibition in anaplastic thyroid cancer and enhancement of ICAM-1-targeted chimeric antigen receptor (CAR)-T cell tumor lysis. Cancer Res. 78:2738. 2018.
50	Williams P, Basu S, Garcia-Manero G, Hourigan CS, Oetjen KA, Cortes JE, Ravandi F, Jabbour EJ, Al-Hamal Z, Konopleva M, et al: The distribution of T cell subsets and the expression of immune checkpoint receptors and ligands in patients with newly diagnosed and relapsed acute myeloid leukemia. Cancer. 125:1470–1481. 2019. View Article : Google Scholar : PubMed/NCBI