Generation of cytotoxic aptamers specifically targeting fibroblast-like synoviocytes by CSCT-SELEX for treatment of rheumatoid arthritis.

OBJECTIVES: Rheumatoid arthritis (RA) is an autoimmune disease characterised by aggressive fibroblast-like synoviocytes (FLSs). Very few RA patients-derived FLSs (RA-FLSs)-specific surface signatures have been identified, and there is currently no approved targeted therapy for RA-FLSs. This study aimed to screen therapeutic aptamers with cell-targeting and cytotoxic properties against RA-FLSs and to uncover the molecular targets and mechanism of action of the screened aptamers. METHODS: We developed a cell-specific and cytotoxic systematic evolution of ligands by exponential enrichment (CSCT-SELEX) method to screen the therapeutic aptamers without prior knowledge of the surface signatures of RA-FLSs. The molecular targets and mechanisms of action of the screened aptamers were determined by pull-down assays and RNA sequencing. The therapeutic efficacy of the screened aptamers was examined in arthritic mouse models. RESULTS: We obtained an aptamer SAPT8 that selectively recognised and killed RA-FLSs. The molecular target of SAPT8 was nucleolin (NCL), a shuttling protein overexpressed on the surface and involved in the tumor-like transformation of RA-FLSs. Mechanistically, SAPT8 interacted with the surface NCL and was internalised to achieve lysosomal degradation of NCL, leading to the upregulation of proapoptotic p53 and downregulation of antiapoptotic B-cell lymphoma 2 (Bcl-2) in RA-FLSs. When administrated systemically to arthritic mice, SAPT8 accumulated in the inflamed FLSs of joints. SAPT8 monotherapy or its combination with tumour necrosis factor (TNF)-targeted biologics was shown to relieve arthritis in mouse models. CONCLUSIONS: CSCT-SELEX could be a promising strategy for developing cell-targeting and cytotoxic aptamers. SAPT8 aptamer selectively ablates RA-FLSs via modulating NCL-p53/Bcl-2 signalling, representing a potential alternative or complementary therapy for RA.

Repurposing AS1411 for constructing ANM-PROTACs.

Proteolysis-targeting chimeras (PROTACs) are heterobifunctional molecules consisting of two ligands joined by a linker, enabling them to simultaneously bind with an E3 ligase and a protein of interest (POI) and trigger proteasomal degradation of the POI. Limitations of PROTAC include lack of potent E3 ligands, poor cell selectivity, and low permeability. AS1411 is an antitumor aptamer specifically recognizing a membrane-nucleus shuttling nucleolin (NCL). Here, we repurpose AS1411 as a ligand for an E3 ligase mouse double minute 2 homolog (MDM2) via anchoring the NCL-MDM2 complex. Then, we construct an AS1411-NCL-MDM2-based PROTAC (ANM-PROTAC) by conjugating AS1411 with large-molecular-weight ligands for "undruggable" oncogenic STAT3, c-Myc, p53-R175H, and AR-V7. We show that the ANM-PROTAC efficiently penetrates tumor cells, recruits MDM2 and degrades the POIs. The ANM-PROTAC achieves tumor-selective distribution and exhibits excellent antitumor activity with no systemic toxicity. This is a PROTAC with built-in tumor-targeting and cell-penetrating capacities.

Association between tumor morphology and dosimetric parameters of organs at risk after intensity-modulated radiotherapy in esophagus cancer.

PURPOSE: We explored the effects of geometrical topological properties of tumors such as tumor length and "axial cross-sectional area (ACSA)" of tumors (planning target volume [PTV] volume /PTV length) on the dosimetric parameters of organs at risk (lung and heart) in patients with esophagus cancer (EPC) treated by way of intensity-modulated radiation therapy (IMRT), so as to provide a guideline for the dosimetric limitation for organs at risk in IMRT treatment. METHODS: A retrospective analysis was done on 103 cases of patients with EPC who were treated by IMRT from November 2010 to August 2019, in which PTV-G stood for the externally expanded planning target volume (PTV) of the gross tumor volume (GTV) and PTV-C for the externally expanded volume of the clinical target volume (CTV). A linear regression model was employed to analyze the several pairs of correlation: the 1st one between the relative length of tumors (PTV length/lung length) and pulmonary dose-volume parameters, the 2nd one between ACSA of tumors and pulmonary dose-volume parameters, the 3rd one between PTV length and the dosimetric parameters of the heart, and the last one between ACSA of tumors and the dosimetric parameters of the heart. RESULTS: (i) There was a strong positive correlation between the relative length of tumors (PTV length/lung length) and V5 (p < 0.001, r = 0.73), and V10 (p < 0.001, r = 0.66) of the lung. There was a moderate positive correlation between the relative length of tumors and V30 (p < 0.001, r = 0.44) of the lung, and a weak positive correlation between the relative length of tumors and V20 (p < 0.001, r = 0.39) of the lung. (ii) There was a strong positive correlation between ACSA of tumors (PTV volume/PTV length) and V30 (p < 0.001, r = 0.67) of the lung, a moderate positive correlation between ACSA of tumors and V20 (p <0.001, r = 0.51) of the lung, and a weak positive correlation between ACSA of tumors and V10 (p = 0.019, r = 0.23) of the lung, yet there was not an obvious correlation between ACSA of tumors and V5 p > 0.05) of the lung. (iii) There was a moderate positive correlation between PTV length and V40 (p < 0.001, r = 0.58), and Dmean (p < 0.001, r = 0.52) of the heart, yet there was no obvious correlation between ACSA of tumors and Dmean and V40 of the heart (p > 0.05). CONCLUSIONS: (i) Compared with the high-dose region of the lung, the relative length of tumors (PTV length/lung length) has a greater impact on the low-dose region of the lung. The linear regression equation of scatter plot showed that when the relative length of tumors increased by 0.1, the lung dose-volume parameters of V5 , V10 , V20 , and V30 increased by approximately 5.37%, 3.59%, 1.05%, and 1.08%, respectively. When PTV length increased by 1 cm, Dmean and V40 of the heart increased by approximately 153.6 cGy and 2.03%, respectively. (ii) Compared with the low-dose region of the lung, the value of ACSA of tumors (PTV volume/PTV length) has a greater impact on the high-dose region of the lung. However, the value of ACSA of tumors has no significant effect on the dosimetric parameters of the heart (Dmean and V40 ). The linear regression equation of scatter plot showed that when ACSA of tumors increased by 10 cm2 , the lung dose-volume parameters of V10 , V20, and V30 increased by approximately 3.11%, 3.37%, and 4.01%, respectively.

The growth feature and its diagnostic value for benign and malignant pulmonary nodules met in routine clinical practice.

BACKGROUND: Growth rate is an independent risk factor for lung cancer in screened pulmonary nodules. This study aimed to clarify growth characteristics of pulmonary nodules in routine clinical practice and examine whether volume doubling time (VDT) can predict the malignancy of these nodules. METHODS: We retrospectively enrolled patients with 5-30-mm-sized pulmonary nodules that had been surgically resected after a follow-up of at least 3 months. Two follow-up computed tomography (CT) images with similar thickness and long interval were obtained. Then, three-dimensional (3D) manual segmentation for all nodules was performed on two follow-up CT scans. Subsequently, VDT was calculated for nodules with a change in volume of at least 25%. RESULTS: Overall, 305 pulmonary nodules in 305 patients (men, 36.7%; median age, 57) were included. The mean increased diameter, mass, and volume of benign (n=86) and malignant (n=219) nodules were 0.09 vs. 2.37 mm, 0.10 vs. 0.66 g, and 32.74 vs. 1,871.28 mm3, respectively (P<0.05). In total, 24 of 86 benign nodules (28%, 18 grew and 6 shrank) and 121 of 219 malignant nodules (55%, 114 grew and 7 shrank) changed over time. The median VDTs of growing benign and malignant nodules were 389 and 526 days, respectively, (P=0.18), and the area under the receiver operating characteristic (ROC) curve was 0.67 (0.55-0.78), with a sensitivity and specificity of 69% and 58%, respectively. The median VDT for growing nodules was 339 days for inflammatory pseudotumors, 226 days for granulomas, 640 days for benign tumors, 1,541 days for enlarged lymph nodes, 762 days for adenocarcinoma in situ, 954 days for microinvasive adenocarcinoma, 534 days for invasive adenocarcinoma, and 118 days for squamous cell carcinoma. CONCLUSIONS: In routine clinical practice, many malignant nodules could grow slowly or even remain stable over time. Regarding growing nodules, the diagnostic value of VDT was limited.

Newly excysted juveniles (NEJs) of Fasciola gigantica induce mice liver fibrosis and M2 macrophage-like phenotype in vivo.

Liver flukes of animals are parasitic flatworms of major socioeconomic importance in many countries. Particularly, Fasciola gigantica is a leading cause of production losses to the livestock (mainly sheep and cattle) and meat industries due to clinical disease, reduced weight gain and milk production, and deaths. Immune responses induced by helminth have been extensively studied, but there is limited information on this aspect by F. gigantica, especially on macrophages induced with this parasite. Studies have shown that host immune responses induced by parasitic infection is greatly correlated with the macrophage polarization axis. In the present study, we used the murine model of F. gigantica to explore the interaction of host and F. gigantica. We found F. gigantica NEJs promoted pathology and fibrosis of mice liver, and the enlargement of mice spleen. We also showed that macrophages were recruited to mice peritoneal cavity at 5 days post infection. By evaluating the expression of genetic markers of M2 macrophages such as Arg-1, Ym1 and RELMɑ, and genetic marker of M1 macrophages iNOS, we showed that M2 macrophages were induced by F. gigantica. M2 macrophages are central to the immune response during helminth infection, and our findings in this study provided insight into the immune interaction between F. gigantica and host.