GAP JUNCTION FUNCTION IS ESSENTIAL FOR SURVIVAL OF ACUTE LYMPHOBLASTIC LEUKEMIA CELLS
DOI:
https://doi.org/10.15407/exp-oncology.2024.02.110Keywords:
gap junction, stromal cells, bone marrow, acute lymphoblastic leukemia, carbenoxolone, methotrexateAbstract
Background. Acute lymphoblastic leukemia has an intimate physical relationship with nonmalignant bone marrow stromal cells. We have recently demonstrated that stromal cells contribute to the survival of leukemia cells and that there is a bidirectional transfer of intracellular material between them. Understanding the mechanisms of stromal support of leukemia may provide insights into new therapies. Aim. To test the hypothesis that gap junctions are formed between acute lymphoblastic leukemia cells and nonmalignant stromal cells, and that gap junction function is essential for the survival of leukemia cells. Materials and Methods. We employed a well-characterized in vitro model of human bone marrow stromal cells and primary human B lymphoblastic leukemia cells and measured leukemia cell survival in coculture using flow cytometry. We measured the effects of gap junction antagonist peptides, carbenoxolone (a drug known to interfere with the gap junction function), and several leukemia chemotherapy drugs including methotrexate upon leukemia cell survival. Results. We demonstrated that stromal cells need to be alive and metabolically active to keep leukemia cells alive. Physical contact between stromal and leukemia cells leads to an increase in gap junction proteins in leukemia cells. Gap junction inhibitory peptides impaired leukemia cell survival as did carbenoxolone, a nonpeptide inhibitor of the gap junction function. Stromal cell survival was not affected. We observed a very modest enhancement of methotrexate antileukemia activity by low-dose carbenoxolone but no significant interactions with dexamethasone, vincristine, mercaptopurine, or doxorubicin. Conclusion. These studies demonstrate that acute lymphoblastic cell survival is impaired by interference with the gap junction function. The development of drugs targeting gap junctions may provide a novel approach to the therapy of acute lymphoblastic leukemia.
References
Usmani S, Sivagnanalingam U, Tkachenko O, et al. Support of acute lymphoblastic leukemia cells by nonmalignant bone marrow stromal cells. Oncol Lett. 2019;17(6):5039-5049. https://doi.org/10.3892/ol.2019.10188
Grosely R, Kopanic JL, Nabors S, et al. Effects of phosphorylation on the structure and backbone dynamics of the intrinsically disordered connexin43 C-terminal domain. J Biol Chem. 2013;288(34):24857-24870. https://doi. org/10.1074/jbc.M113.454389
Kim MO, Lee YJ, Han HJ. Involvement of Cx43 phosphorylation in 5'-N-ethylcarboxamide-induced migration and proliferation of mouse embryonic stem cells. J Cell Physiol. 2010;224(1):187-194. https://doi.org/10.1002/jcp.22117
Gonzalez-Nieto D, Li L, Kohler A, et al. Connexin-43 in the osteogenic BM niche regulates its cellular composition and the bidirectional traffic of hematopoietic stem cells and progenitors. Blood. 2012;119(22):5144-5154. https://doi. org/10.1182/blood-2011-07-368506
Dürig J, Rosenthal C, Halfmeyer K, et al. Intercellular communication between bone marrow stromal cells and CD34+ haematopoietic progenitor cells is mediated by connexin 43-type gap junctions. Br J Haematol. 2000;111(2):416-425. https://doi.org/10.1046/j.1365-2141.2000.02385.x
Rosendaal M, Krenács TT. Regulatory pathways in blood-forming tissue with particular reference to gap junctional communication. Pathol Oncol Res. 2000;6(4):243-249. https://doi.org10.1007/bf03187326
Cancelas JA, Koevoet WL, de Koning AE, et al. Connexin-43 gap junctions are involved in multiconnexin-expressing stromal support of hemopoietic progenitors and stem cells. Blood. 2000;96(2):498-505. PMID: 10887111
Rosendaal M, Green CR, Rahman A, Morgan D. Up-regulation of the connexin43+ gap junction network in haemo- poietic tissue before the growth of stem cells. J Cell Sci. 1994;107(Pt 1):29-37. https://doi.org/10.1242/jcs.107.1.29
Singh AK, Cancelas JA. Gap junctions in the bone marrow lympho-hematopoietic stem cell niche, leukemia progres- sion, and chemoresistance. Int J Mol Sci. 2020;21(3). https://doi.org/10.3390/ijms21030796
Yang S, Wen Q, Liu Y, et al. Increased expression of CX43 on stromal cells promotes leukemia apoptosis. Oncotarget. 2015;6(42):44323-44331. https://doi.org/10.18632/oncotarget.6249
Zhang X, Liu Y, Si YJ, et al. Effect of Cx43 gene-modified leukemic bone marrow stromal cells on the regulation of Jurkat cell line in vitro. Leuk Res. 2012;36(2):198-204. https://doi.org/10.1016/j.leukres.2011.10.001
Mihara K, Imai C, Coustan-Smith E, et al. Development and functional characterization of human bone marrow mesenchymal cells immortalized by enforced expression of telomerase. Br J Haematol. 2003;120(5):846-849. https:// doi.org/10.1046/j.1365-2141.2003.04217.x
Rajnai H, Teleki I, Kiszner G, et al. Connexin 43 communication channels in follicular dendritic cell development and in follicular lymphomas. J Immunol Res. 2015;2015:528098. https://doi.org/10.1155/2015/528098
Tarzemany R, Jiang G, Larjava H, Hakkinen L. Expression and function of connexin 43 in human gingival wound healing and fibroblasts. PloS One. 2015;10(1):e0115524. https://doi.org/10.1371/journal.pone.0115524
Moore AR, Zhou WL, Sirois CL, et al. Connexin hemichannels contribute to spontaneous electrical activity in the hu- man fetal cortex. Proc Natl Acad Sci U S A. 2014;111(37):E3919-E3928. https://doi.org/10.1073/pnas.1405253111
Pinder RM, Brogden RN, Sawyer PR, et al. Carbenoxolone: a review of its pharmacological properties and therapeu- tic efficacy in peptic ulcer disease. Drugs. 1976;11(4):245-307. https://doi.org/10.2165/00003495-197611040-00002
Downer HD, Galloway RW, Horwich L, Parke DV. The absorption and excretion of carbenoxolone in man. J Pharm Pharmacol. 1970;22(7):479-487. https://doi.org/10.1111/j.2042-7158.1970.tb10550.x
Kouzi F, Zibara K, Bourgeais J, et al. Disruption of gap junctions attenuates acute myeloid leukemia chemoresistance induced by bone marrow mesenchymal stromal cells. Oncogene. 2020;39(6):1198-1212. https://doi.org/10.1038/ s41388-019-1069-y
Sinyuk M, Alvarado AG, Nesmiyanov P, et al. Cx25 contributes to leukemia cell communication and chemosensiti- vity. Oncotarget. 2015;6(31):31508-31521. https://doi.org/10.18632/oncotarget.5226
Tittarelli A, Guerrero I, Tempio F, et al. Overexpression of connexin 43 reduces melanoma proliferative and meta- static capacity. Br J Cancer. 2015;113(2):259-267. https://doi.org/10.1038/bjc.2015.162
Chen Q, Boire A, Jin X, et al. Carcinoma-astrocyte gap junctions promote brain metastasis by cGAMP transfer. Na- ture. 2016;533(7604):493-498. https://doi.org/10.1038/nature18268
Uzu M, Sato H, Yamada R, Kashiba T, Shibata Y, Yamaura K, Ueno K. Effect of enhanced expression of connexin 43 on sunitinib-induced cytotoxicity in mesothelioma cells. J Pharmacol Sci. 2015;128(1):17-26. https://doi.org/10.1016/j. jphs.2015.04.002
Huang T, Zhu Y, Fang X, et al. Gap junctions sensitize cancer cells to proteasome inhibitor MG132-induced apopto- sis. Cancer Sci. 2010;101(3):713-721. https://doi.org/10.1111/j.1349-7006.2009.01421.x
Jensen R, Glazer PM. Cell-interdependent cisplatin killing by Ku/DNA-dependent protein kinase signaling transduced through gap junctions. Proc Natl Acad Sci U S A. 2004;101(16):6134-6139. https://doi.org/10.1073/pnas.0400051101
Carystinos GD, Alaoui-Jamali MA, Phipps J, et al. Upregulation of gap junctional intercellular communication and connexin 43 expression by cyclic-AMP and all-trans-retinoic acid is associated with glutathione depletion and chemosensitivity in neuroblastoma cells. Cancer Chemother Pharmacol. 2001;47(2):126-132. https://doi. org/10.1007/s002800000231
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2024 Experimental Oncology
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
