ObjectiveChronic kidney disease (CKD) is a major global health problem. In clinical practice, the Chinese patent herbal medicine Jianpi-Yishen (JPYS) formula is commonly used to treat CKD. However, the molecular mechanisms by which JPYS targets and modulates the host immune response remain unclear.MethodsThis study utilized network pharmacology, RNA sequencing (RNA-seq), and metabolic analyses using in vivo and in vitro models to investigate the impact of the JPYS formula on inflammation and the immune system. Specifically, the study focused on macrophage polarization and metabolic changes that may slow down the progression of CKD.ResultsA total of 14,946 CKD-related targets were identified from the GeneCards and Online Mendelian Inheritance in Man (OMIM) databases through network pharmacology analyses. 227 potential targets of the JPYS formula were predicted using the TCMSP database. Additionally, network diagram demonstrated that 11 targets were associated with macrophage activity. In vivo studies indicated that the JPYS formula could reduce blood urea nitrogen and serum creatinine in adenine-induced CKD rats. Furthermore, the formula inhibited inflammatory damage and abnormal macrophage infiltration in this CKD model. RNA-seq, proteomic and metabolic analyses identified the regulation of amino acid metabolism by betaine, specifically referring to glycine, serine, and threonine metabolism, as a key target of the JPYS formula in slowing the progression of CKD. In addition, in vitro studies suggested that JPYS may enhance tryptophan metabolism in M1 macrophage polarization and betaine metabolism in M2 macrophage polarization.ConclusionsThe JPYS formula has been shown to have beneficial impact on CKD; a key mechanism is the mitigation of inflammatory damage through the interaction between amino acid metabolism and macrophage polarization. Of specific importance in this context are the roles of tryptophan in M1 polarization and betaine in M2 polarization.

Introduction Triptolide (TPL) is a promising plant-derived compound for clinical therapy of multiple human diseases; however, its application was limited considering its toxicity. Methods To explore the underlying molecular mechanism of TPL nephrotoxicity, a network pharmacology based approach was utilized to predict candidate targets related with TPL toxicity, followed by deep RNA-seq analysis to characterize the features of three transcriptional elements include protein coding genes (PCGs), long noncoding RNAs (lncRNAs) and circular RNAs (circRNAs) as well as their associations with nephrotoxicity in rats with TPL treatment. Results & Discussion Although the deeper mechanisms of TPL nephrotoxcity remain further exploration, our results suggested that c-Jun is a potential target of TPL and Per1 related circadian rhythm signaling is involved in TPL induced renal toxicity.

Females typically outlive males, a disparity mitigated by castration, yet the molecular underpinnings remain elusive. Our study integrates multi-omics and behavioral analyses to uncover the pivotal compounds and genes influencing healthy aging post-castration, examining serum, kidney, and liver biospecimens from 12-week and 18-month old castrated male mice and their unaltered counterparts. Behavioral tests and LC-MS/MS metabolomics reveal that castrated males exhibit altered steroid hormones, superior cognitive performance, and higher levels of anti-oxidative compounds like taurine, despite identical diets. Integrated metabolome-transcriptome analysis confirms reduced lipid peroxidation and oxidative stress in female and castrated male mice, suggesting a protective mechanism against aging. Histological examinations post-cisplatin treatment highlight the model’s applicability in studying sex-dependent drug toxicity and reveal varying susceptibility in organ-specific toxicities, underlining the crucial role of sex hormones in physiological defenses. In essence, our castration model unveils a feminized metabolic and transcriptomic intermediary, serving as a robust tool for studying sex-specific aspects of healthy aging and exploring sex hormone-induced differences in diverse biomedical domains.### Competing Interest StatementThe authors have declared no competing interest.

Background. Danggui-Shaoyao-San (DSS) is a traditional Chinese medicine formula that has been widely used to treat a variety of disorders, including renal diseases. Despite being well-established in clinical practice, the mechanisms behind the therapeutic effects of DSS on diabetic nephropathy (DN) remain elusive. Methods. To explore the therapeutic mechanism, we explored the action mechanism of DSS on DN using network pharmacology strategies. All ingredients were selected from the relevant databases, and active ingredients were chosen on the basis of their oral bioavailability prediction and drug-likeness evaluation. The putative proteins of DSS were obtained from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, whereas the potential genes of DN were obtained from the GeneCards and OMIM databases. Enrichment analysis using gene ontology (GO) and the Kyoto encyclopedia of genes and genomes (KEGG) was performed to discover possible hub targets and gene-related pathways. Afterwards, the underlying molecular mechanisms of DSS against DN were validated experimentally in vivo against db/db mice. Results. We identified 91 phytochemicals using the comprehensive network pharmacology technique, 51 of which were chosen as bioactive components. There were 40 proteins and 20 pathways in the target-pathway network. The experimental validation results demonstrated that DSS may reduce the expression of TNF-α, IL-6, and ICAM-1, as well as extracellular matrix deposition, by blocking the JNK pathway activation, which protects against kidney injury. Conclusion. This study discovered the putative molecular mechanisms of action of DSS against diabetic kidney damage through a network pharmacology approach and experimental validation.

杨曙东教授认为慢性肾脏病的病机以脾肾阳虚为本,湿浊、瘀血壅扰为标,治疗应以温法为主,再根据患者舌、脉、症状变化,辅以清、消、补等法.针对脾肾阳虚,治以辛热扶阳、甘温补气,处方常用金匮肾气丸、补中益气汤、四君子汤等;针对寒湿表证,治以温阳散寒,处方常用麻黄加白术汤、白术附子汤、桂枝附子汤等;针对水湿痰瘀,治以温清并用,根据三焦病位选择相应药物;针对瘀血,治以温阳化瘀,处方多以黄芪桂枝五物汤、桃红四物汤加减.附验案一则.

Objective: Based on the ancient and modern medical record cloud platform, to analyze the medication rule of Dai medicine Maoxucao in treatment of chronic kidney disease.Methods: Collect the medical record data of chronic kidney disease treated with felwort, extract the medical record information that meets the inclusion criteria from the outpatient system, and establish the prescription database.Use the methods of statistical analysis, cluster analysis, complex network analysis and other methods of system integration to analyze the frequency of prescription and drug compatibility of the medical record.Results: 149 medical records were finally included, and the analysis indicates that the commonly used ingredients with high compatibility are Baizhu（atractylodes macrocephala）,Niuxi（achyranthes bidentata）,Duzhong（eucommia ulmoides oliv）,etc.The drug properties are mainly mild, warm, and slightly cold, and the taste are mainly sweet, bitter, and sour, most of which attributed to the Liver, Spleen and Kidney meridians.The drug efficacy is to strengthen the Liver and Kidney and to strengthen the muscles and bones.Three results of drug cluster analysis were obtained; The core prescription drugs commonly used for compatibility obtained from complex network analysis are Baizhu（atractylodes macrocephala）,Niuxi（achyranthes bidentata）,Duzhong（eucommia ulmoides oliv）,Taizishen（radix Pseudostellariae）,Guijianyu（ramuli euonymi）,Baishao（radix paeoniae alba）,Mudanpi（peony bark）,Maidong（ophiopogon japonicus） and Mohanlian（Eclipta）;Further analysis of the relationship between "prescription and medicine" and "symptom and medicine" leads to the rule of drug compatibility for the treatment of foam urine, edema, fatigue and other symptoms.Conclusion: In the clinical application of spicate clerodendranthus herb in treating chronic kidney disease, we should focus on the treatment based on syndrome differentiation and pay attention to the combined use of multiple drugs so as to strengthen the Zheng Qi and remove the evil factors.

Background. Fatigue is a common symptom in adults that may cause physical and psychological problems and reduce quality of life. Aromatherapy could possibly provide relief for those suffering from fatigue. Here, we evaluated the effect of aromatherapy on fatigue in adults. Methods. We searched the PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure, Chinese Biomedical Literature, SinoMed, Wanfang, and Chinese Scientific Journal Database databases for randomized controlled trials of aromatherapy treatment for fatigue in adults from their inception to June 2021. Two reviewers searched independently, extracted the characteristics of the studies, and assessed the risk of bias using the Cochrane risk-of-bias tool and Stata v. 14.0. Results. Nineteen studies were included in this systematic review. Aromatherapy had a significant effect on fatigue (standardized mean difference −0.64, 95% confidence interval−1.14, −0.15, I2 94.4%, P<0.001). Subgroup analysis according to aromatic type, substance, frequency, treatment duration, intervention, outcomes measurement, and population type showed that aromatherapy had a significantly greater effect in the intervention group, compared to the control group. Funnel plots and Egger’s test indicated no significant publication bias. Conclusion. Our results suggest that aromatherapy ameliorates fatigue in adults who suffer from chronic diseases. A rigorous intervention program and larger randomized controlled trials are needed.

李顺民教授承袭国医大师邓铁涛"五脏相关"理论并结合多年临床经验,提出从"脾肾相关"治疗肾性蛋白尿的思路.认为肾性蛋白尿的病机特点为脾肾亏虚为本,兼以湿热、湿浊、瘀血、风邪扰肾为标;治疗以健脾补肾为治疗总则,兼顾利湿、清热、祛风,并在治疗的全过程注重调畅气血.健脾可选补中益气汤加减,补肾常用参芪地黄汤加减.通过调理脾肾脏腑气血,可协调五脏,保护肾脏.

Platelet-derived growth factor (PDGF) signaling, besides other growth factor-mediated signaling pathways like vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), seems to play a crucial role in tumor development and progression. We have recently provided evidence for upregulation of PDGF expression in UICC stage I-IV primary colorectal cancer (CRC) and demonstrated PDGF-mediated induction of PI3K/Akt/mTOR signaling in CRC cell lines. The present study sought to follow up on our previous findings and explore the alternative receptor cross-binding potential of PDGF in CRC. Our analysis of primary human colon tumor samples demonstrated upregulation of the PDGFRβ, VEGFR1, and VEGFR2 genes in UICC stage I-III tumors. Immunohistological analysis revealed co-expression of PDGF and its putative cross-binding partners, VEGFR2 and EGFR. We then analyzed several CRC cell lines for PDGFRα, PDGFRβ, VEGFR1, and VEGFR2 protein expression and found these receptors to be variably expressed amongst the investigated cell lines. Interestingly, whereas Caco-2 and SW480 cells showed expression of all analyzed receptors, HT29 cells expressed only VEGFR1 and VEGFR2. However, stimulation of HT29 cells with PDGF resulted in upregulation of VEGFR1 and VEGFR2 expression despite the absence of PDGFR expression and mimicked the effect of VEGF stimulation. Moreover, PDGF recovered HT29 cell proliferation under simultaneous treatment with a VEGFR or EGFR inhibitor. Our results provide some of the first evidence for PDGF cross-signaling through alternative receptors in colorectal cancer and support anti-PDGF therapy as a combination strategy alongside VEGF and EGF targeting even in tumors lacking PDGFR expression.

透析相关性低血压是导致透析患者终点事件发生的独立危险因素之一,严重影响患者生活.西医现有的治疗手段给患者带来的长期影响还尚未明确,而中医结合理、法、方、药等独特的辨证论治体系在临床治疗此病时取得了显著疗效.此文从中医学角度对透析相关性低血压的文献古籍研究、现代医家观点进行阐述,并结合临床研究,从病因病机、辨证施治、药物作用机制等方面,多方面整理研究了此病的中医药治疗现状.

Clear cell renal cell carcinoma (ccRCC) is the most lethal form of kidney cancer and effective treatment regimens are yet to be established. Tyrosine kinase inhibitors (TKI) have widely been used as ccRCC therapeutics, but their efficacy is limited due to accompanying resistance mechanisms. Previous studies have provided substantial evidence for crosstalk between cAMP and the MAPK/ERK signaling pathway. Low levels of intracellular cAMP have been found in several human malignancies and some data suggest that elevation of cAMP expression can be achieved by phosphodiesterase 4 (PDE4) inhibition, resulting in cell growth arrest and/or cell death. The effects of crosstalk between cAMP and the MAPK/ERK pathway on the development progression in ccRCR, however, remain to be fully understood. In this study, we sought to explore the involvement of PDE4 in ccRCC and to assess its potential as a target for therapeutic intervention. We demonstrated that PDE4D is the predominant subtype of PDE4 expressed in healthy and cancerous renal cell lines, particularly in metastatic Caki-1 cells. We generated a CRISPR/Cas9-mediated PDE4D-KO Caki-1 cell model and showed that PDE4D depletion reduced cell proliferation and recovered cAMP expression in these cells. PDE4D-KO and/or PDE4 inhibition with the FDA approved PDE4 inhibitor, roflumilast, also attenuated MAPK/ERK signaling in a CRAF-dependent manner. Most interestingly, we showed that PDE4D-KO enhanced the effectiveness of the TKI, sorafenib, to stunt cell survival. In conclusion, we provide preliminary evidence of PDE4 involvement in ccRCC and suggest a rationale for dual tyrosine kinase/PDE4D targeting in patients with CRAF-dependent MAPK activation.

Background Non-high-density lipoprotein cholesterol (non-HDL-C) may be an independent risk factor for cardio-cerebrovascular disease (CVD); however, the cutoff level in patients on maintenance hemodialysis (MHD) is unknown. Methods This was a retrospective multicenter study of MHD patients treated at 10 dialysis centers in Guangdong Province from July 1, 2016, to April 1, 2017. Laboratory test data were collected and CVD complications and outcomes recorded. Results In total, 1288 eligible patients were included in this study; the non-HDL-C interquartile range was 2.76 (2.24-3.45) mmol/L. Over a median follow-up time of 24 months, 141 patients developed CVD. The non-HDL-C level was a principal risk factor for such events (P < 0.05; 95% confidence interval 0.800-0.842). The maximum Youden index was 0.549 and the best cutoff > 3.39 mmol/L. Conclusion Higher baseline non-HDL-C levels may increase the CVD risk in MHD patients. Thus, non-HDL-C effectively predicts CVD.

Abstract Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest types of cancer with 5-year survival rates between 5-8%. Chronic inflammation of the pancreas and pancreatitis is thought to be a risk factor in PDAC. The aim of this study was to elucidate characteristic pro-inflammatory cytokines and chemokines including IL-17 profiles in human pancreatic cancer. A total of 186 individuals with confirmed PDAC (n=116), chronic pancreatitis (n=32), and healthy volunteers (n=38) were included in the study. The histological stage of the tumor was determined according to the Union for International Cancer Control (UICC) TNM staging system. Tumors were evaluated for stage and differentiation grade. Data concerning age, gender, level of wall infiltration, lymph node metastasis, and distant metastases were collected in a database. Additional clinical characteristics necessary for statistical analysis of the data from the study population were collected from the Tumor Registry. Inflammation-related cytokines and chemokines in IL-17 pathway among the patients were analyzed from serum obtained pre-operatively and compared with healthy controls. Moreover, the KEGG PATHWAY, a collection of manually drawn pathway maps on molecular interaction, reaction and relation networks was used for IL-17 pathway analysis. Non-parametric tests and correlation tests were utilized to compare the factors among three groups for statistical analysis. Pancreatitis patients showed highest expression in inflammatory cytokines and chemokine panels while those with PDAC showed different profiles. Pro-inflammatory cytokines including IL-5, IL-6, CXCL8, CCL2, TNF-alpha, and IFN-gamma revealed higher expression in PDAC patients than healthy volunteers (P<0.05). Anti-inflammatory cytokines like IL-13 demonstrated lower expression in PDAC patients than healthy controls (P<0.05), which was likewise observed for IL-17. Interestingly, PDAC patients with distant metastases showed lower pro-inflammatory expression profiles than those with localized tumors. Moreover, correlation analysis demonstrated that IL-17 was correlated with IL-13 and GM-CSF expression (P<0.05). In conclusion, we provide evidence that PDAC patients are characterized by dominant expression of pro-inflammatory cytokine profiles, diminished IL-17 level within their serum, and also anti-inflammatory mediators within is in part divergent from other solid cancers. The IL-17 signaling. may therefore play a different role in the inflammatory pathway in pancreatic cancer. Citation Format: Yueming Luo, Martin Gasser, Amrendra Ajay, Li-LI Hsiao, Ana Maria Waaga-Gasser. Pro-inflammatory serum marker profiles that lack upregulated IL-17 are characteristic for pancreatic cancer and require for further anti-tumor immune response and strategies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1758.

Background. Coronavirus disease 2019 (COVID-19) causes psychological distress and can have a negative impact on the general mental health and rehabilitation in affected patients under currently implemented isolation guidelines. Auricular point pressure (APP) as well-established technique in traditional Chinese medicine may help to relieve sleep disturbance and anxiety in COVID-19 patients. Methods. During the early phase of the epidemic/pandemic, patients were enrolled in this study (02/2020 until 03/2020 n = 84). They were strictly isolated on specific wards at the Hubei Provincial Hospital of Integrated Chinese and Western Medicine in Hubei. The retrospective cohort study design included two groups. Group A patients were treated with an auricular point pressure (APP) in addition to standard intensive care medicine while Group B participants (No-APP) received routine nursing measures alone. Treatment outcome was measured using the St. Mary’s Hospital Sleep Questionnaire (SMH) Score and the 7-Item Generalized Anxiety Disorder Scale (GAD-7). Both scores were measured in each patient at baseline and on the discharge day. Results. The SMH score and sleep status changed in APP patients at the end of the treatment period when compared with No-APP patients ( P < 0.01 ). APP-treated patients demonstrated lower GAD-7 scores than No-APP controls ( P < 0.01 ). Further, no significant differences in safety or adverse events between the APP and No-APP groups were observed. Conclusion. The results from our snapshot study during the early phase of the SARS-CoV-2 epidemic/pandemic suggest that auricular point pressure could be a simple and effective tool to relieve insomnia and situational anxiety in hospitalized patients suffering from COVID-19 and kept under disconcerting conditions of isolation.

Abstract Anti-Epithelial/Vascular Endothelial Growth Factor (EGF/VEGF)-targeted therapy plays important roles in advanced stage colorectal cancer (CRC) patients but fails to show clinical efficacy in specific subgroups which lack of clear underlying mechanisms. In vitro studies indicated that cross-talk or alternative receptor/ligand activities can bypass anti-growth factor signaling arrest and therefore may explain failure of monoclonal antibody (mAb) targeted therapy. In this study we analyzed effects of Platelet Derived Growth Factor (PDGF)-mediated bypassed signaling in CRC. We first investigated human UICC (Union for International Cancer Control ) stage I-IV tumors (n=46) from our Center and found increased PDGFRβ and VEGFR1/2 gene expression (p<0.01/p<0.001). Interestingly, further analysis on human HT29 colon cancer in vitro showed upregulated VEGFR1/2 expression upon VEGF stimulation. This was even more pronounced upon stimulation with PDGF. Inhibition of VEGFR2 and EGFR resulted in decreased HT29 cell proliferation, while additional treatment with VEGF and particularly PDGF attenuated the inhibiting effect and resulted in altered downstream signaling events. Treatment with the tyrosine kinase inhibitor Regorafenib alone, that blocks VEGFR/EGFR+PDGFR, prevented PDGF and VEGF-induced proliferation. This effect was not observed for either VEGFR2 mAb treatment with Ramucirumab or EGFR mAb therapy with Cetuximab. To conclude, PDGF-induced human colon cancer proliferation in the absence of PDGFR and simultaneously inhibited VEGFR2 and EGFR expression in HT29 colon cancer cells suggests bypassed alternative PDGF receptor partners for ligand binding on the tumor cell surface and subsequent intracellular signaling. This may explain failure of anti-VEGF/EGF-targeted monoclonal antibody therapies in CRC patient subgroups and underlines the necessity to effectively inhibit PDGF-mediated signaling events beside VEGF/EGF blockade in patients with increased PDGF expression within their tumors. Citation Format: Romana Moench, Martin Gasser, Karol Nawalaniec, Tanja Grimmig, Minghua Cao, Yueming Luo, Li-Li Hsiao, Ana Maria Waaga-Gasser. PDGF-induced tumor cell growth despite VEGF/EGF inhibition and absence of PDGF receptor expression in colon cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3755.

BACKGROUND:The application of factor analysis in the study of the clinical symptoms of coronavirus disease 2019 (COVID-19) was investigated, to provide a reference for basic research on COVID-19 and its prevention and control.METHODS:The data of 60 patients with COVID-19 in Jingzhou Hospital of Traditional Chinese Medicine and the Second People's Hospital of Longgang District in Shenzhen were extracted using principal component analysis. Factor analysis was used to investigate the factors related to symptoms of COVID-19. Based on the combination of factors, the clinical types of the factors were defined according to our professional knowledge. Factor loadings were calculated, and pairwise correlation analysis of symptoms was performed.RESULTS:Factor analysis showed that the clinical symptoms of COVID-19 cases could be divided into respiratory-digestive, neurological, cough-wheezing, upper respiratory, and digestive symptoms. Pairwise correlation analysis showed that there were a total of eight pairs of symptoms: fever-palpitation, coughexpectoration, expectoration-wheezing, dry mouth-bitter taste in the mouth, poor appetite-fatigue, fatiguedizziness, diarrhea-palpitation, and dizziness-headache.CONCLUSIONS:The symptoms and syndromes of COVID-19 are complex. Respiratory symptoms dominate, and digestive symptoms are also present. Factor analysis is suitable for studying the characteristics of the clinical symptoms of COVID-19, providing a new idea for the comprehensive analysis of clinical symptoms.

Introduction: Clinical studies have indicated a relationship between diabetic nephropathy (DN) and the incidence and prevalence of renal cell carcinoma (RCC). However, the mechanism linking diabetic nephropathy and renal cell carcinoma has not yet to be identified. Methods: In this study, a total of 42 male Sprague Dawley (SD) rats were randomly assigned to a DN group (n=35) and a control group (n=7). All animals in the DN group were unilaterally nephrectomized and treated with streptozotocin with the development of blood glucose levels >16.7mmol/L and dominant proteinuria and were compared to controls without such changes. Histopathologic alterations in the kidneys were examined by HE staining and Ki-67 immunohistochemistry. Differentially expressed genes were identified and validated by RNA-seq and PCR. Results: As the results, except for two rats that failed to develop the DN model and were excluded from the analysis, 33 rats in the DN group with overt signs of DN demonstrated significantly higher food and water intake, urine production, and urine protein and urinary protein/creatinine ratio than controls. Overall, 15.2% (n=5/33) of DN animals developed RCC while none tumors were observed in the control group (n=0/7). RNA-seq analysis in these animals indicated different TRPV5 gene expression and calcium path-way expression in DN animals with developing tumors, when compared with animals with no obvious tumors. In addition, DN animals diagnosed with RCC showed increased expression of GLUT2 and c-met, when compared to controls and DN animals without tumors. Discussion: In conclusion, the disordered calcium metabolism, especially disturbed TRPV5 mediated Ca2+ signal, may have been related to the development of RCC in DN rats. Further studies related to the detailed mechanism are still needed.

目的 通过知识图谱挖掘技术分析杨霓芝教授益气活血治疗糖尿病肾病的临证思路,为临床治疗提供指导.方法 系统收集广东省中医院杨霓芝教授门诊2007年5月至2017年7月治疗糖尿病肾病病历,严格筛选后通过广东省中医院大数据团队中医药大数据智能处理与知识服务系统进行相关数据挖掘,并对专家进行访谈,转换为书面文字整理.结果 杨霓芝教授擅长使用益气活血法治疗糖尿病肾病,在遣方用药方面有律可循,主要以黄芪、白术、菟丝子、山萸肉、太子参等益气补肾药和丹参、桃仁、泽兰、当归等活血药为主;太子参-旱莲草、丹参-黄芪、当归-淫羊藿为常用药对;大黄为糖尿病肾病后期慢性肾衰的专药.以益气活血法为主线,可以根据糖尿病肾病不同阶段进行化裁.结论 本研究对益气活血法的中药、配伍及药-症关系加以综述研究,表明杨霓芝教授治疗糖尿病肾病以益气活血法为主,配伍得当,药症关系密切,为今后进一步的临床研究提供思路与依据.

1Nephrology Department, State Key Laboratory of Dampness Syndrome of Chinese Medicine, Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of China; 2Department of Nephrology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen Traditional Chinese Medicine Hospital, Shenzhen, Guangdong, People’s Republic of China; 3Transplantation Research Center, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA; 4Department of Pathology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of China; 5Department of Pediatrics, Guangdong Second Hospital of Chinese Medicine, Guangzhou, People’s Republic of China

Abstract Clear cell renal cell carcinoma (ccRCC) is the most lethal form of kidney cancer. Small molecule VEGFR inhibitors are widely used but are not curative and various resistance mechanisms have been described. Previous studies have indicated cross-talk between the cAMP pathway and the MAPK pathway, and the elevation of cAMP levels by inhibition of Phosphodiesterase-4 (PDE4) activities can result in growth arrest and/or cell death. In this study we focused on the effects of PDE4 in ccRCC. We first analyzed the expression of PDE4 in human RCC cell lines and found PDE4D to be dominantly expressed. PDE4D CRISPR Knockout (KO) was then performed on Caki-1 RCC cells and normal renal proximal tubular epithelial cells as control. Cell proliferation, cell cycle cAMP, MAPK plus PI3K/AKT signaling pathways were additionally analyzed to define the effects of PDE4D inhibition. PDE4D KO Caki-1 cells showed decreased cell viability and a higher apoptosis rate compared with wild type controls. Increased intracellular cAMP levels in PDE4D KO resulted in downregulated MAPK but not PI3K/AKT signaling in Caki-1 tumor cells. In addition, PDE4D KO Caki-1 cells demonstrated higher sensitivity to VEGFR inhibitors. To conclude, we provide a new preclinical rationale for dual PDE4/VEGFR2 inhibition in patients with ccRCC. While the MAPK signaling pathway is activated in ccRCC, dual inhibitors may improve the anti-tumor effects in a patient subset with evidence of MAPK involvement. Future work involving in vivo models will be useful to better define efficacy of this dual strategy of anti-tumor activity. Citation Format: Minghua Cao, Karol Nawalaniec, Yueming Luo, Romana Moench, Li-Li Hsiao, Ana Maria Waaga-Gasser. Combined PDE4 and VEGFR2 inhibition as a new preclinical rationale in clear cell renal cell carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6386.