Clanxet et al. BMC Veterinary Research (2022) 18:206 https://doi.org/10.1186/s12917-022-03317-1 RESEARCH Open Access Gene expression profiles of beta‑adrenergic receptors in canine vascular tumors: a preliminary study Jordi Clanxet1, Mariana Teles2,3 , Javier Hernández‑Losa4 , Manuel Ruiz‑Echarri Rueda5, Luis Benitez‑Fusté6 and Josep Pastor1*    Abstract Beta adrenergic receptors (β-AR) play a key role in regulating several hallmark pathways of both benign and malig‑ nant human and canine tumors. There is scarce information on the expression of β-AR in canine vascular tumors. Therefore, the purpose of the present research work was to study the mRNA expression levels of the three subtypes of the β-AR genes (ADRB1, ADRB2, ADRB3) in hemangiosarcoma (HSA) and hemangioma (HA), as well as in vascular hamartomas (VH) from dogs. Fifty samples (n = 50) were obtained from 38 dogs. Twenty-three animals had HSA, eight animals HA and seven animals VH. HSA were auricular (n = 8), splenic (n = 5), cutaneous (n = 6), auricular and splenic (n = 2), cutaneous- muscular (n = 1) and disseminated (n = 1). There were seven cases of HSA that were divided into primary tumor and secondary (metastatic) tumor. Skin and muscle samples with a normal histological study were used as control group. ADRB gene expression was determinate in all samples by real-time quantitative PCR. Results showed that ADRB1, ADRB2 and ADRB3 were overexpressed in HSA when compared to the control group. ADRB2 was overexpressed in HA when compared to the control group. HSA express higher values of ADBR1 (p = 0.0178) compared to VH. There was a high inter-individual variability in the expression of the three subtypes of ADBR. No statistically significant difference in the expression of ADBR genes were observed between HSA primary when compared to metastatic or in different anatomical locations. In conclusion, canine HSA overexpress the three β-AR subtypes and canine HA β2-AR. High vari‑ ability was observed in β-AR mRNA levels amongst HSA cases. Keywords: ADBR, Beta receptors, Dog, Hamartoma, Hemangiosarcoma, Hemangioma Introduction subtype is mainly located in the heart, kidney and adi- Beta adrenergic receptors (β-AR) play a key role in pocytes; β2-AR is mainly distributed in the lungs and regulating several hallmark pathways of both benign bronchi, vascular smooth muscle, sympathetic terminal, and malignant human and canine tumors [1–4]. There heart, uterus, skeletal muscle, among others; and β3-AR are three subtypes of β-AR (β1, β2 and β3) with a vari- is found in adipocytes, uterus, bladder and heart [1, 2]. able physiological distribution in tissues. The β1-AR The three subtypes of β-AR have also been identified in primary and metastasis tumors of the brain, lung, liver, kidney, adrenal glands, ovary, prostate, lymphoid tissue, *Correspondence: josep.pastor@uab.cat melanoma, bone marrow and vasculature [5, 6]. 1 In human medicine, the β1-AR and β2-AR subtypes Department of Animal Medicine and Surgery, Universitat Autònoma de Barcelona, 08193 Barcelona, Spain are involved in tumoural mechanisms, such as prolifera- Full list of author information is available at the end of the article tion and angiogenesis. Recently, the β3-AR subtype has © The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://​creat​iveco​mmons.​org/​licen​ses/​by/4.​0/. The Creative Commons Public Domain Dedication waiver (http://​creat​iveco​ mmons.​org/​publi​cdoma​in/​zero/1.​0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Clanxet et al. BMC Veterinary Research (2022) 18:206 Page 2 of 7 also been found to take part in these pro-tumoral pro- selected. All dogs were sampled with the owner’s per- cesses [7]. Moreover, β2-AR has been specially described mission for clinical or diagnostic reasons, and an owner as overexpressed in human angiosarcoma. Furthermore, informed consent was obtained in order to use those recent data revealed a strong correlation between the use samples for future studies. Inclusion criteria was a for- of β-AR antagonists and a significant reduction of the malin-fixed and paraffin embedded tissue sample with a cancer progression, metastasis and mortality in affected histological diagnosis of HA, HSA or VH. Both primary patients, even in advanced stages of the disease [4, 8, 9]. and metastatic HSA were included in the study. Primary Also the therapeutic targeting of the β-AR has shown and secondary tumors were classified based on the clini- efficacy in the treatment of benign vascular tumors, such cal record and followed up of the case, in cases where as infantile hemangiomas, with propranolol becoming followed up were not available the larger tumor was con- the treatment of election since its serendipitous discov- sidered the primary specially if they affect the auricle or ery [10, 11]. Thus, β-AR antagonists could be used as an the spleen As a control group, skin and muscle samples adjunct drug to current therapeutic strategies in clinical with a normal histological diagnosis were used. Con- oncology [5, 9, 12, 13]. trol samples were processed in the same way that tumor Vascular tumors are a common neoplasia in dogs, samples. being hemangioma (HA, benign) and hemangiosarcoma (HSA, malignant) the most frequent vascular tumors RNA extraction, cDNA synthesis and β‑adrenergic diagnosed [14, 15]. Canine HA are mostly located in the receptors (ADRB) gene expression skin (84%) and it is widely distributed in the head, neck, Only paraffin conserved samples with more than 800% of trunk and extremities. HA account for approximately tumor sample confirmed by histology were used. Paraffin 4.5% of all skin neoplasms and HSA for less than 1% [16, conserved tissue samples were numerated from 1 to 50 17]. HA has a benign behavior and complete surgical in order to have each case identified. For some animals resection is usually curative [15]. Canine HSA is a highly a pool of tumors in different localization was used to aggressive neoplasia that is usually visceral [18]. This type extract RNA as it is stated in table 1 at the supplemen- of tumor is characterized by a high metastatic rate and a tary material). After deparaffinization of 10 µm paraffin poor response to the current available treatment options, embedded tissue samples, RNA was extracted using the which leads to a poor prognosis [4, 19]. Surgery is the High Pure FFPET RNA Isolation Kit (Roche). Cells were treatment of choice for most primary HSA. However, due lysed in 100 µL RNA Tissue Lysis Buffer, 16 µL 10% SDS to its highly aggressive behavior and the poor outcome of and 40 µL of Proteinase K during 30 min at 85 ºC and the surgery alone, adjuvant chemotherapy is necessary shaking at 600 rpm. After this step, 60 µL of Proteinase K in all cases [4, 19]. HSA have been presumed to have its were added and the tissue samples were incubated at 55 origin from transformed endothelial cells, although, new ºC and shaking at 600 rpm for 30 min. data suggest a pluripotent bone marrow progenitor as the After cell lysis, the samples were incubated during cell of origin [20]. 15 min at 15 to 25 ºC with the DNAase working solution Hamartomas are unifocal/multifocal overgrowths of to enhance the RNA extraction. After this, the samples mature non-tumoral cells and tissue native to the organ were centrifuged several times to eliminate debris and, in which it occurs. Due to their limited growth, they are finally, 50 µL of RNA Elution Buffer was added in order considered developmental malformations [21]. Vascular to obtain the pure total RNA. RNA quantification quan- hamartomas (VH) are the most frequent form of hamar- tity (ng/µL) and the quality (260/280 ratio) was done tomas found in domestic animals [22] and correspond to using a NanoDrop Spectrophotometer (Thermo Fisher a disorganized and excessive proliferation of the vascular Scientific). tissue [23]. Reverse transcription (RT) was performed with 1 μg of With all the above in mind, the purpose of the present RNA using the High-Capacity cDNA Reverse Transcrip- research work was to study the mRNA expression levels tion Kit (Thermo Fisher Scientific, ABI Applied Biosys- of the three subtypes of the β-AR genes (ADRB1, ADRB2, tems 9902 Veriti PCR Thermal Cycler). The program was ADRB3) in canine vascular tumors (HA and HSA), as 10 min at 25 ºC, 120 min at 37 ºC, 5 min at 85 ºC and well as in VH. forever at 4 ºC. For the ADRB gene expression RT-qPCR was per- Materials and methods formed using the following primers: ADRB1 (Biorad; Criteria for case inclusion Dog #ENSCAFT00000017826.4), ADRB2 (Biorad; Samples from biopsies and necropsies from the Diag- Dog #ENSCAFT00000029135.3), ADRB3 (Biorad; nostic Service of Veterinary Pathology of the Universi- Dog #ENSCAFT00000009965.3), SDHA (Biorad; Dog tat Autònoma de Barcelona (UAB) were retrospectively #ENSCAFT00000017502.3) and GAPDH (Biorad; Dog Clanxet et al. BMC Veterinary Research (2022) 18:206 Page 3 of 7 #ENSCAFT00000023939.3). SDHA and GAPDH were Most animals were over eight years old in the two groups Each well contained 10 µL of 2 × SsoAdvanced™ Uni- selected as housekeeping genes. of animals with types of tumors and in the group of ani- versal SYBR® Green, 1 µL of 20 × PrimePCR, 8 µL of mals with the vascular malformation (n = 27; 71%). The mean age of the animals for each condition was: 8.2 years nuclease-free water and 1 µL of cDNA. Control samples, old for VH, 8.7 years old for HA and 9.7 years old for nuclease-free water and six cDNA samples were included HSA. There were, eight animals with HA, twenty-three in triplicate for each primer in all the plates. animals with HSA and seven animals with VH. HSA were After assessing primers assembly, RT-qPCR was per- auricular (n = 8), splenic (n = 5), cutaneous (n = 6), auric- formed using the 7500 Fast Real-Time PCR System ular and splenic (n = 2), cutaneous-muscular (n = 1) and (Applied Biosystems) for the ADRB gene expression. The disseminated (n = 1). There were seven cases of HSA that RT-qPCR program was: 2 min at 95 ºC (1 cycle) and 5 s at were divided into primary tumor and secondary (meta- 95 ºC and 30 s at 60 ºC (40 cycles). Gene expression levels static) tumor because both samples were analyzed. VH were determined using the ΔΔCt method normalized to were located in encephalon (n = 2), scrotum (n = 2), tes- the ­Ct mean of GAPDH and SDHA. Skin and muscle tis- ticle (n = 1) and periosteum (n = 1). HA were cutaneous sue samples from twelve healthy dogs were used as a con- (n = 5), subcutaneous (n = 2) and epithelioid (n = 1). trol group and the three subtypes of ADRB expression was expressed as fold changes in VH, HA and HSA. Fold ADRB1, ADRB2 and ADRB3 expression in HA, HSA and VH changes were calculated with the formula ­2−ΔΔCt. Figure 1 shows the mRNA levels of the 3 ADRB in HA, HSA and VH. ADRB1 (p = 0.0011) and ADRB3 Statistical analysis (p = 0.0046) were overexpressed in HSA groups when The expression of ADBR genes amongst vascular tumors compared to the control group, while ADRB2 was over- were studied using a non -parametric ANOVA test with expressed in HA (p = 0.0305) and HSA (p = 0.0020) when Dunn’s multiple comparison test. Mann Whitney test compared to the control group. HSA express statisti- was used to compare ADBR expression in primary and cally significantly higher values of ADBR1 (p = 0.0178) secondary HSA. Finally, we studied the ADBR expres- compared to VH. Note that there is a high inter-indi- sion depending on HSA tissue localization with Kruskall- vidual variability in the expression of the three subtypes Wallis test. Significance was set at p < 0.05 in all cases and of ADBR. However, the levels of gene expression of the tests were carried out with GraphPad Prism 8 (GraphPad three ADRB show a clear tendency to increase with the Software, San Diego, CA, USA). malignancy of the tumor. Results Tissue samples ADRB1, ADRB2 and ADRB3 gene expression in primary Fifty samples (n = 50) were obtained from 38 dogs, and secondary HSA twenty-four were males and fourteen were females (Sup- We also studied if the expression levels of the three plementary material Table 1). The most affected breeds ADRB subtypes was different in primary HSA and sec- were German Shepherd (one HA, nine HSA and one VH), ondary (metastatic) HSA. We did not find statistically Crossbreed (two HA and six HSA), Boxer (three HA and significant differences in the expression levels of the one VH) and Labrador Retriever (one HA and one HSA). ADRB1 (p = 0,6761), ADRB2 (p = 0,5244) and ADRB3 Fig. 1 Gene expression of the 3 β-adrenergic receptors subtypes in tissues of control animals and animals presenting hemangiomas (HA), hemangiosarcomas (HSA) and vascular hamartomas (VH). * (p < 0.05) and ** (p < 0.001) means significant different between groups Clanxet et al. BMC Veterinary Research (2022) 18:206 Page 4 of 7 (p = 0,3157) subtypes between primary and metastatic of cancer [4]. β1-AR overexpression has been demon- tumors (Fig. 2). strated in human HSA [9, 13], while β2-AR overexpres- sion is variable [9]. Another study affirms that β2-AR ADRB1, ADRB2 and ADRB3 gene expression depending is highly overexpressed in human and canine HSA cell on the tissue localization of the HSA lines [27]. β3-AR is poorly expressed in human HSA, We also studied whether the expression levels of the three unlike canine HSA cell lines, which predominantly over- ADBR changed with the HSA tumor localization. We did express β2-AR and β3-AR [27]. In a more recent study not find statistically significant differences in the expres- on 10 samples from visceral HSA and 5 non-malignant sion levels of any of the three ADRB subtypes: ADRB1 splenic hematomas, HSA appeared to express higher lev- (p = 0.8803), ADRB2 (p = 0,0.9520), ADRB3 (p = 0.7744) els of immunohistochemistry reactivity towards all three when comparing among them (Fig. 3). β-AR when compared to the non-malignant hematoma samples. Also, they located the immunoreactivity against Discussion β1-AR at the nucleus, β2-AR and β3-AR were mainly The canine samples used in the present study come observed in the cytoplasm of tumor cells [28]. In the pre- from a population of animals similar in age and breeds sent study we observed that all 3 β-AR were expressed, described in other studies of canine HSA, HA and VA but the degree of expression was very variable among [10, 16–18, 22–25]. Also, the localization of the primary individual samples. Additionally, we found that they tumor and the metastatic diseases is in concordance with were overexpressed in the primary as well as in second- previous studies and the described progression of canine ary or metastatic disease. However, there are more pri- HSA [10, 17, 18, 22, 26]. Thus, conclusion obtained in mary samples with clear overexpression of β-AR than our study represent the clinical situation of most vas- those considered secondary. This overexpression could cular tumors and malformations observed in veterinary make these samples outliers, the meaning of these outli- medicine. ers is not clear because the overexpression of the β-AR is In humans, the three subtypes of β-AR have been a characteristic of a more malignant phenotype and ini- identified in HA [6] and HSA [6, 13] among other types tially we hypothesized that metastatic samples may have Fig. 2 Gene expression of the 3 β-adrenergic receptors subtypes in primary and secondary (metastatic) hemangiosarcoma (HSA). No statistically significant differences were found Fig. 3 Gene expression of the 3 β-adrenergic receptors subtypes in hemangiosarcoma (HSA) at different localizations. No statistically significant differences were found Clanxet et al. BMC Veterinary Research (2022) 18:206 Page 5 of 7 higher overexpression. Anyway, this observation was not a higher sensitivity to β-AR antagonists against these statistically significant and further studies are needed tumors [27]. Also, in human melanoma and colon can- in order to assess any differences in β-AR expression cer there are reports pointing a promising use of specific between primary and metastatic HSA in the dog.. A gen- betta blockers in their treatment [29, 33–35]. Recently, eral overview of beta adrenergic signalling and the regu- the role of β1-AR in an animal model of cavernous vas- lation of tumor cell biology in the context of human and cular malformations and the effect of its specific block- canine HSA have been published [4]. One of the limita- ing in preventing these malformations has been studied tions of our study is the lack of follow up for most of the [36]. In our study, we could not demonstrate a clear over- cases studied. It will be interesting to document if β-AR expression of β-AR receptors in VA and the impact of expression is correlated not only with the malignant phe- β-adrenergic blocker in reducing vascular malformations notype but also with prognosis or responds to standard is not clearly predicted as it has been proved in this ani- chemotherapy treatment. Another limitation of our study mal model and human medicine (31). is that we used skin and muscle as control, and they have In conclusion, this study showed overexpression of the low expression of β-AR, probably VH could be better three β-AR subtypes in canine HSA and β2-AR in canine control, however, using be this strategy we ensure that HA. High variability was observed in β-AR mRNA levels we assessed uniformly the β-AR expression in all vascular amongst HSA cases, and we theorize that β-AR antago- tumors without losing the data of the VH. nists would be useful in dogs with a high β-AR overex- The main interest in the expression of β-AR in tumoral pression. Further research is needed to understand the tissues is to evaluate the potential effects of the use of role of β-AR in canine vascular tumors, specially HSA, blocker therapy in preventing tumors proliferation [29]. and the efficacy of β-AR blockers therapy. It is clear that in human medicine, propranolol is used as a frontline therapy in the treatment of infantile HA since Supplementary Information 2015 [30] and it has been proposed for the treatment of The online version contains supplementary material available at https://​doi.​ other vascular abnormalities [31]. In the current study, org/​10.​1186/​s12917-​022-​03317-1. HA significantly overexpressed the ADBR2 gene (β2-AR), Additional file 1: Table S1. Summary of canine samples studied with thus it can be hypothesized that a similar clinical situa- breed, gender, histological diagnosis and tumor localization from where tion can be expected to happen in canine patients with samples were obtained. HA. However, future prospective studies will be needed to demonstrate that hypothesis. Acknowledgements Propranolol, a β-AR antagonist, has been used with Mariana Teles acknowledge the Spanish Government (Ministerio de Ciencia e Innovación) for her contract with reference RYC2019-026841-I. The promising outcomes in the treatment of human HSA [4, authors also acknowledge Joan Carles Balasch for the graphical design of the 9–11]. In our study, and in previous studies with canine figures. HSA cells, it has been demonstrated that all three β-AR Authors’ contributions are overexpressed [23, 28]. Thus, it can also be hypoth- JC, JP, LB, JHL and MRR collected and analyzed the presented results. JC, esized that its blocking can improve tumor control and MT, JP wrote the main manuscript text and prepared Figs. 1, 2 and 3. All prolong survival. Also, a recent study suggests that pro- authors reviewed the manuscript. The author(s) read and approved the final manuscript. pranolol synergizes with doxorubicin by a decrease lyso- somal sequestration and cellular efflux of doxorubicin Funding while increasing its intracellular concentration and con- The authors have no disclaimers or source of support to disclose. sequently apoptosis [28]. However, as it can be seen in Availability of data and materials our study results, the expression of β-AR is very vari- Material available by request to the corresponding authors (josep.pastor@uab. able among individual tumors and the impact of the use cat). of β-AR antagonist needs to be addressed in future pro- spective studies and in animals were β-AR is documented Declarations by inmmunohistochemistry or qPCR in order to obtain Ethics approval and consent to participate adequate data to unravel the β-AR role in canine HSA. During this study no animal experimentation has been done, and only left- Although, there are controversy in human medicine and over used for other diagnostic purposes has been used. For this, all dogs were sampled with the owner’s permission for clinical or diagnostic reasons, and an some reports in human breast cancer and neuroblastoma owner informed consent was obtained in order to use the left-over in future did not show a clear correlation between β-AR mRNA studies. By doing this we followed the recommended guidelines of the Ethical levels and increased susceptibility to β-AR antagonist committee of the Universitat Autònoma de Barcelona and Hospital Clinic Veterinary and also we followed relevant guidelines as the Directive 2010/63/ therapy [6, 12, 32]. On the other hand, other publica- EU in Europe). tions demonstrated high levels of mRNA expression in malignant vascular tumors and, therefore, they suggested Clanxet et al. BMC Veterinary Research (2022) 18:206 Page 6 of 7 Consent for publication 14. Chikweto A, McNeil P, Bhaiyat MI, Stone D, Sharma RN. Neoplastic and Not aplicable. nonneoplastic cutaneous tumors of dogs in Grenada. West Indies Int Sch Res Not. 2011;2011:1–6. https://​doi.​org/​10.​5402/​2011/​416435. Competing interests 15. Corbally MT. Soft-tissue sarcoma. In: Small Animal Clinical Oncology. 5th The authors declare that they have no known competing financial interests ed. Amsterdam: Elsevier; 2013. p. 930–4. or personal relationships that could have appeared to influence the work 16. Gross TL, Ihrke PJ, Walder EJ, Affolter VK. “Vascular tumors.” In: Skin Dis‑ reported in this paper. eases of the Dog and Cat. 2nd ed. Oxford: Blackwell Science Ltd; 2005. p. 735–58. Author details 17. Gamlem H, Nordstoga K, Arnesen K. Canine vascular neoplasia - a 1 Department of Animal Medicine and Surgery, Universitat Autònoma de population-based clinicopathologic study of 439 tumours and tumour- Barcelona, 08193 Barcelona, Spain. 2 Department of Cell Biology, Physiology like lesions in 420 dogs. APMIS. 2009;116(125):41–54. https://​doi.​org/​10.​ and Immunology, Universitat Autònoma de Barcelona, 08193 Barcelona, 1111/j.​1600-​0463.​2008.​125m4.x. Spain. 3 Institute of Biotechnology and Biomedicine, Universitat Autònoma de 18. Schultheiss PC. A retrospective study of visceral and nonvisceral heman‑ Barcelona, 08193 Barcelona, Spain. 4 Molecular Biology Laboratory, Depart‑ giosarcoma and hemangiomas in domestic animals. J Vet Diagnostic ment of Pathology, Hospital Universitario Vall d’Hebron, Passeig Vall d´Hebron, Investig. 2004;16(6):522–6. https://​doi.​org/​10.​1177/​10406​38704​01600​606. 119‑129, 08035 Barcelona, Spain. 5 Bristol Royal Infirmary, Marlborough Street, 19. Mullin C, Clifford CA. Miscellaneous tumors: hemangiosarcoma. In: Vail Bristol BS2 8HW, UK. 6 Hospital Veterinari del Mar, C/ Marina 69, 08005 Barce‑ DM, Thamm DH, Liptak JM, editors. Withrow & MacEwen’s Small Animal lona, Spain. Clinical Oncology. 6th ed. St Louis: Elsevier; 2020. p. 773–8. 20. Kim J, Graef AJ, Dickerson EB, Modiano JF. Pathobiology of hemangio‑ Received: 23 February 2022 Accepted: 18 May 2022 sarcoma in dogs: research Advances and future perspectives. Vet Sci. 2015;2(4):388–405. https://​doi.​org/​10.​3390/​vetsc​i2040​388. 21. Shankargouda P, Roopa SR, Majumdar B. Hamartomas of the oral cavity. J Int Soc Prev Community Dent. 2015;5(5):347–53. https://​doi.​org/​10.​4103/​ 2231-​0762.​164789. References 22. Gualtieri M, Cocci A, Monti S, Olivero D. Surgical removal of a localised 1. Daly CJ, Mcgrath JC. Previously unsuspected widespread cellular and tis‑ vascular hepatic hamartoma in a dog. Aust Vet J. 2009;87(9):360–2. sue distribution of beta-adrenoceptors and its relevance to drug action. https://​doi.​org/​10.​1111/j.​1751-​0813.​2009.​00472.x. Trends Pharmacol Sci. 2011;32(4):219–26. https://​doi.​org/​10.​1016/j.​tips.​ 23. Sakurai M, Morita T, Kondo H, Uemura T, Haruna A, Shimada A. Cer‑ 2011.​02.​008. ebral vascular hamartoma with thrombosis in a dog. J Vet Med Sci. 2. Stein CM. “β-Adrenergic Receptors.” In: Primer on the Autonomic Nervous 2011;73(10):1367–9. https://​doi.​org/​10.​1292/​jvms.​11-​0201. System. 3rd ed. Amsterdam: Elsevier; 2012. p. 59–61. 24. Yasuno K, Kobayashi R, Ohmuro T, Kamiie J, Sahara H, Shirota K. Caudal 3. Coelho M, Soares-Silva C, Brandao D, Marino F, Cosentino M, Ribeiro vascular hamartoma accompanied by aberrant arteriovenous structures L. Beta-adrenergic modulation of cancer cell proliferation: Avail‑ in a dog. J Vet Diagnostic Investig. 2011;23(5):1051–5. https://​doi.​org/​10.​ able evidence and clinical perspectives. J Cancer Res Clin Oncol. 1177/​10406​38711​416852. 2017;143(2):275–91. https://​doi.​org/​10.​1007/​s00432-​016-​2278-1. 25. Smith SH, Van Winkle T. Cerebral vascular hamartomas in five dogs. Vet 4. Dickerson E, Bryan B. Beta adrenergic signalling: A targetable regulator of Pathol. 2001;38(1):108–12. https://​doi.​org/​10.​1354/​vp.​38-1-​108. angiosarcoma and hemangiosarcoma. Vet Sci. 2015;2(3):270–92. https://​ 26. Moe L, Gamlem H, Dahl K, Glattre E. Canine neoplasia - population-based doi.​org/​10.​3390/​vetsc​i2030​270. incidence of vascular tumours. APMIS. 2008;125:63–8. https://​doi.​org/​10.​ 5. Cole SW, Sood AK. Molecular pathways: beta-adrenergic signalling in 1111/j.​1600-​0463.​2008.​125m6.x. cancer. Clin Cancer Res. 2012;18(5):1201–6. https://​doi.​org/​10.​1158/​1078-​ 27. Stiles JM, Amaya C, Rains S, Diaz D, Pham R, Battiste J, et al. Targeting 0432.​CCR-​11-​0641. of beta adrenergic receptors results in therapeutic efficacy against 6. Rains SL, Amaya CN, Bryan BA. Beta-adrenergic receptors are expressed models of hemangioendothelioma and angiosarcoma. PLoS One. across diverse cancers. Oncoscience. 2017;4(7–8):12–5. https://​doi.​org/​10.​ 2013;8(3):e60021. https://​doi.​org/​10.​1371/​journ​al.​pone.​00600​21. 18632/​oncos​cience.​357. 28. Saha J, Kim JH, Amaya CN, Witcher C, Khammanivong A, Korpela DM, 7. Escarcega González CE, Rodríguez MG, Marichal Cancino BA, González et al. Propranolol sensitizes vascular sarcoma cells to doxorubicin by Hernández A, Villalón C. β-adrenoceptor blockade for infantile altering lysosomal drug sequestration and drug efflux. Front Oncol. hemangioma therapy: Do β3-adrenoceptors play a role? J Vasc Res. 2021;10(2):1–15. https://​doi.​org/​10.​3389/​fonc.​2020.​614288. 2018;55(3):159–68. https://​doi.​org/​10.​1159/​00048​9956. 29. Filippi L, Bruno G, Domazetovic V, Favre C, Calvani M. Current thera‑ 8. Amaya CN, Perkins M, Belmont A, Herrera C, Vargas A, Khayou T, et al. pies and new targets to fight melanoma: a promising role for the Non-selective beta blockers inhibit angiosarcoma cell viability and β3-adrenoreceptor. Cancers. 2020;12(6):1–16. https://​doi.​org/​10.​3390/​ increase progression free- and overall-survival in patients diagnosed with cance​rs120​61415. metastatic angiosarcoma. Oncoscience. 2018;5(3–4):109–19. https://​doi.​ 30. Léauté-Labrèze C, Hoeger P, Mazereeuw-Hautier J, Guibaud L, Baselga org/​10.​18632/​oncos​cience.​413. E, Posiunas G, et al. A randomized, controlled trial of oral propranolol in 9. Pasquier E, André N, Street J, Chougule A, Rekhi B, Ghosh J, et al. Effective infantile hemangioma. N Engl J Med. 2015;372(8):735–46. https://​doi.​org/​ management of advanced angiosarcoma by the synergistic combina‑ 10.​1056/​nejmo​a1404​710. tion of propranolol and vinblastine-based metronomic chemotherapy: a 31. Trenor C. Medical management of vascular anomalies. Semin Cutan Med bench to bedside study. EBioMedicine. 2016;6:87–95. https://​doi.​org/​10.​ Surg. 2016;35(3):177–81. https://​doi.​org/​10.​12788/j.​sder.​2016.​046. 1016/j.​ebiom.​2016.​02.​026. 32. Wolter JK, Wolter NE, Blanch A, Partridge T, Cheng L, Morgenstern DA, 10. Sans V, de la Roque ED, Berge J, Grenier N, Boralevi F, Mazereeuw-Hautier et al. Anti-tumor activity of the beta-adrenergic receptor antagonist J, et al. Propranolol for severe infantile hemangiomas: follow-up report. propranolol in neuroblastoma. Oncotarget. 2014;5(1):161–72. https://​doi.​ Pediatrics. 2009;124(3):423–31. https://​doi.​org/​10.​1542/​peds.​2008-​3458. org/​10.​18632/​oncot​arget.​1083. 11. Léauté-Labrèze C, de la Dumas RE, Hubiche T. Propranolol for severe 33. Calvani M, Pelon F, Comito G, Taddei ML, Moretti S, Innocenti S, et al. hemangiomas of infancy. N Engl J Med. 2008;358(24):2649–52. https://​ Norepinephrine promotes tumor microenvironment reactivity through doi.​org/​10.​1056/​NEJMc​07088​19. β3-adrenoreceptors during melanoma progression. Oncotarget. 12. Chisholm KM, Chang KW, Truong MT, Kwok S, West RB, Heerema-Mcken‑ 2015;6(7):4615–32. https://​doi.​org/​10.​18632/​oncot​arget.​2652. ney AE. β-Adrenergic receptor expression in vascular tumors. Mod Pathol. 34. Perrone MG, Notarnicola M, Caruso MG, Tutino V, Scilimati A. Upregula‑ 2012;25(11):1446–51. https://​doi.​org/​10.​1038/​modpa​thol.​2012.​108. tion of β3-adrenergic receptor mRNA in human colon cancer: a prelimi‑ 13. Chow W, Amaya CN, Rains S, Chow M, Dickerson EB, Bryan BA. Growth nary study. Oncology. 2008;75(3–4):224–9. https://​doi.​org/​10.​1159/​00016​ attenuation of cutaneous angiosarcoma with propranolol-mediated 3851. β-blockade. JAMA Dermatol. 2015;151(11):1226–9. https://​doi.​org/​10.​ 35. Montoya A, Amaya CN, Belmont A, Diab N, Trevino R, Villanueva G, 1001/​jamad​ermat​ol.​2015.​2554. et al. Use of non-selective β-blockers is associated with decreased Clanxet et al. BMC Veterinary Research (2022) 18:206 Page 7 of 7 tumor proliferative indices in early stage breast cancer. Oncotarget. 2017;8(4):6446–60. https://​doi.​org/​10.​18632/​oncot​arget.​14119. 36. Li W, Shenkar R, Detter MR, Moore T, Benavides C, Lightle R, et al. Propran‑ olol inhibits cavernous vascular malformations by β1 adrenergic receptor antagonism in animal models. J Clin Invest. 2021;131(3):e144893. https://​ doi.​org/​10.​1172/​JCI14​4893. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in pub‑ lished maps and institutional affiliations. Ready to submit your research ? Choose BMC and benefit from: • fast, convenient online submission • thorough peer review by experienced researchers in your field • rapid publication on acceptance • support for research data, including large and complex data types • gold Open Access which fosters wider collaboration and increased citations • maximum visibility for your research: over 100M website views per year At BMC, research is always in progress. Learn more biomedcentral.com/submissions