The effect of erythropoietin on normal and neoplastic cells

Figures & data

Figure 1 Erythropoiesis and the expression of stage-specific markers.

Notes: Shown is a schematic diagram of the various stages of human erythropoiesis that results in the formation of mature red blood cells (RBCs). Time-dependent expression of various proteins is shown, including erythropoietin receptor (EpoR) and other cell surface markers (green), transcription factors (orange), and effector molecules such as hemoglobin (red).

Figure 2 (A and B) Erythropoietin (Epo) mRNA is expressed in kidney interstitial cells. Mice were made anemic by withdrawing 0.5 mL blood and replacing with 0.5 mL saline 8, 16, and 24 hours prior to sacrifice. Standard in situ hybridization (ISH) on kidney sections was performed with an antisense ³³P-labeled Epo probe. (A) ISH for mouse Epo mRNA in a control mouse; (B) ISH for mouse Epo mRNA in an anemic mouse.

Notes: Increased number of cells expressing Epo transcripts in kidney from anemic mice vs normal mice, but with a similar number of grains over renal Epo-producing cells from both normal and anemic kidneys. Data and figure kindly provided by Sheilah Scully, Amgen.

Figure 3 Erythropoietin receptor (EPOR), GATA-1, and SCL/Tal1 have similar transcript profiles in normal human tissue.

Notes: Illustrated are levels of transcript (average of n = 2) and standard error obtained through microarray analysis of normal human tissue for EPOR, GATA-1, and SCL/ Tal1. Levels of expression are in mean fluorescence units. Data were obtained from the publicly available database http://biogps.org. Probes shown are: EPOR, 209962_AT; GATA-1, 210446_AT; SCL, 206283-s_AT. Similar intensities were observed with other probes. Note that high-level expression of EPOR mRNA is found primarily in tissue/cell types containing erythroid cells. CD105 (endoglin) is expressed in endothelial cells, but it is also coexpressed with CD71 in erythroid cells.^343,344,461 Thus the EPOR detected in CD105⁺ cells is likely due to erythroid cell–specific expression.

Figure 4 High-level erythropoietin receptor (EpoR) protein expression is found in erythroid cells but not in other tissues. EpoR expression was analyzed by Western immunoblot analysis with anti-EpoR antibody A82 that was shown to specifically detect human EpoR in erythroid cells.⁷⁸ The arrow shows the location of full-length EpoR. Smaller proteins have been shown elsewhere to be EpoR fragments.⁷⁸ UT-7/ Epo cells (EpoR positive control) are derived from a megakaryoblastic leukemia and are Epo-dependent.⁴⁶²

Notes: This research was originally published in Blood. Sinclair AM, Coxon A, McCaffery I, et al. Functional erythropoietin receptor is undetectable in endothelial, cardiac, neuronal, and renal cells. Blood. 2010;115(21):4264–4272. © American Society of Hematology.⁹⁴
Abbreviations: HUVEC, human umbilical vein endothelial cells; RPTEC, renal proximal tubule epithelial cells.

Figure 5 Erythropoietin receptor (EpoR) expression in differentiating CD34+ hematopoietic progenitor cells grown with or without Epo.

Notes: CD34+ cells were cultured in medium containing 50 ng/mL stem cell factor, 10 ng/mL interleukin (IL)-3, 10 ng/mL IL-6 with (w/) and without (no) 5 U/mL recombinant human erythropoietin (rHuEpo) for the indicated number of days. The arrow denotes the position of full-length EpoR (59 kDa). FLAG-EpoR COS7 is an EpoR positive control cell lysate from COS-7 cells expressing a FLAG-tagged version of EpoR.⁷⁶ Data and figure kindly provided by Leigh Busse, Amgen.
Abbreviation: MWM, molecular weight marker in kilo Daltons (kDa).

Figure 6 Erythropoietin receptor (EpoR) activation and signaling with Epo in erythroid progenitor cells.

Note: Schematic diagram of the signaling cascades and effector responses observed in erythroid progenitor cells when EpoR is activated with erythropoiesis-stimulating agents.

Table 1 Effect of erythropoiesis-stimulating agents in xenograft or syngenic tumor models

Tumor type and origin	EPO/DA dose	Reported tumor and survival outcomes	Study
Tumor regression alone
Murine myelomas MOPC-315, 5T33 MM	30 U Epo QD	Tumor regression and prolonged survival	Mittelman et al^Citation440
Murine BCL-1 leukemia/lymphoma	30 U Epo QD	Tumor regression and prolonged survival	Mittelman^Citation441
Enhanced tumor ablation
Neurogenic sarcoma ENE2	750 U/kg Epo TIW	Improved RT therapy in anemic mice	Stuben et al^Citation442
Lewis lung carcinoma	60 U/kg Epo (two doses)	No effect alone; enhanced CT	Sigounas et al^Citation443
Rat DS-sarcoma	1000 U/kg Epo	No effect alone; improved ablative RT	Thews et al^Citation444
Ovary adenocarcinoma	20 U Epo TIW	No effect alone; improved CT	Silver and Piver^Citation445
Glioblastoma HTZ II	1000 U/kg Epo TIW	No effect alone; improved RT in anemic mice	Stuben et al^Citation446
Rat DS-sarcoma	1000 U/kg Epo TIW	No effect alone; improved CT in anemic rats	Thews et al^Citation447
Colon adenocarcinoma	1000 U/kg Epo QD	Restored PT in anemic mice	Golab et al^Citation448
Human glioblastomas GBM-Nan1 and U87	300 U/kg Epo QD	No effect on tumor alone; enhanced RT in both lines	Pinel et al^Citation357
Murine SCC VI squamous cell carcinoma and RIF-1 fibrosarcoma	30 μg/kg DA QW or Q2W	No effect alone; improved RT in anemic mice in both lines	Ning et al^Citation231
Lewis lung carcinoma	10 μg/kg DA QW	No effect alone; improved CT	Shannon et al^Citation270
Human squamous cell A431, colorectal carcinoma HT25	150 U/kg Epo TIW	No effect alone; enhanced CT in both models	Tovari et al^Citation449
Rat breast cancer line LCM 2388 into rat	60 IU Epo QW ± tamoxifen	No effect Epo alone; increased regression Epo + tamoxifen	Sairah et al^Citation356
Human squamous cell carcinoma (A431)	150 U/kg Epo TIW	No effect alone; improved RT in anemic mice	Lovey et al^Citation450
No enhanced tumor ablation
Murine MmB16 melanoma	20 U Epo BID	No effect alone; no enhanced IL-12 therapy	Golab et al^Citation451
Rat R3230 mammary carcinoma	2,000 U/kg Epo TIW	No effect alone	Blackwell et al^Citation452
Rat 13762 mammary adenocarcinoma	50 μg/kg Epo TIW	No effect alone	Bianchi et al^Citation453
Rat DS-sarcoma	1000 U/kg Epo TIW	No effect	Kelleher et al^Citation454
Rat R3230 mammary carcinoma	3 μg/kg DA TIW	No effect to enhance RT	Kirkpatrick et al^Citation455
Murine C26-B colon adenocarcinoma	25 U Epo QD to 25 U TIW	No effect on tumor; decreased body weight loss	van Halteren et al^Citation456
Rat R3230 mammary carcinoma, murine CT26 colon carcinoma, human HCT-116 colon carcinoma, human FaDu head and neck carcinoma	2000 U/kg Epo TIW	No effect alone	Hardee et al^Citation355
Rat R3230 mammary carcinoma	2000 U/kg Epo TIW	No effect alone; no enhanced CT	Hardee et al^Citation457
Human breast carcinomas MDA-MB-231 and MCF-7	2.5 mg/kg epoetin-α, 7.5 mg/kg DA, and 2.5 mg/kg epoetin-β	No effect alone; no enhanced CT in either model	LaMontagne et al^Citation300
Head and neck squamous cell carcinoma LU-HNxSCX-7	400 U/kg epoetin-β Q3D	No effect alone; slight increased tumor growth with surgical transection	Kjellen et al^Citation458
Human breast MCF-7, renal 786-0, gastric SCH, lung A549, ovary SK-OV-3 tumor cell lines into mice	1000, 3000, or 10,000 IU/kg epoetin-β QW	No effect alone; no enhanced effect on bevacizumab on A549 and MCF-7 (avastin)	Kataoka et al^Citation223
Murine B16F10 melanoma	30 mg/kg DA QW	No effect alone	Miller et al^Citation307
Human glioblastoma U87	5000 U/kg Epo TIW	No effect alone	Hassouna et al^Citation299
Human breast MDA453β and MCF7-HER18 (engineered)	100 U rHuEpo daily (weekdays)	No effect alone; antagonized trastuzumab effect on tumor regression	Liang et al^Citation275
Promoted tumor growth
Murine MCA-induced fibrosarcoma	100 IU/kg Epo QW	Epo promoted tumor growth	Okazaki et al^Citation459
Murine colorectal cancer cells in 50% hepatectomized mice	10 mg/kg DA once	Increased tumor growth after hepatectomy	Rupertus et al^Citation460

Abbreviations: TIW, three times per week; BID, twice per day; QD, once daily; QW, once per week; Q2W, every two weeks; Q3D, every three days; RT, radiotherapy; CT, chemotherapy; PT, photodynamic therapy; DA, darbepoetin alfa; Epo, erythropoietin.

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