Transplantation of beige adipose organoids fabricated using adipose acellular matrix hydrogel improves metabolic dysfunction in high-fat diet-induced obesity and type 2 diabetes mice.

Transplantation of brown adipose tissue (BAT) is a promising approach for treating obesity and metabolic disorders. However, obtaining sufficient amounts of functional BAT or brown adipocytes for transplantation remains a major challenge. In this study, we developed a hydrogel that combining adipose acellular matrix (AAM) and GelMA and HAMA that can be adjusted for stiffness by modulating the duration of light-crosslinking. We used human white adipose tissue-derived microvascular fragments to create beige adipose organoids (BAO) that were encapsulated in either a soft or stiff AAM hydrogel. We found that BAOs cultivated in AAM hydrogels with high stiffness demonstrated increased metabolic activity and upregulation of thermogenesis-related genes. When transplanted into obese and type 2 diabetes mice, the HFD + BAO group showed sustained improvements in metabolic rate, resulting in significant weight loss and decreased blood glucose levels. Furthermore, the mice showed a marked reduction in nonalcoholic liver steatosis, indicating improved liver function. In contrast, transplantation of 2D-cultured beige adipocytes failed to produce these beneficial effects. Our findings demonstrate the feasibility of fabricating beige adipose organoids in vitro and administering them by injection, which may represent a promising therapeutic approach for obesity and diabetes.

Transplantation of in vitro prefabricated adipose organoids attenuates skin fibrosis by restoring subcutaneous fat and inducing dermal adipogenesis.

Localized scleroderma is a complex autoimmune disease characterized by dermal fibrosis and loss of cutaneous fat. While cytotherapy offers a promising treatment option, stem cell transplantation results in low survival rates and fails in target cell differentiation. In this study, we aimed to prefabricate syngeneic adipose organoids (ad-organoids) using microvascular fragments (MVFs) via three-dimensional (3D) culturing and transplant them beneath the fibrotic skin to restore subcutaneous fat and reverse the pathological manifestation of localized scleroderma. We employed 3D culturing of syngeneic MVFs with stepwise angiogenic and adipogenic induction to produce ad-organoids and evaluated their microstructure and paracrine function in vitro. C57/BL6 mice with induced skin scleroderma were treated with adipose-derived stem cells (ASCs), adipocytes, ad-organoids, and Matrigel, and the therapeutic effect was assessed histologically. Our results showed that ad-organoids derived from MVF contained mature adipocytes and a well-established vessel network, secreted multiple adipokines, promoted adipogenic differentiation of ASCs, and suppressed proliferation and migration of scleroderma fibroblasts. Subcutaneous transplantation of ad-organoids reconstructed the subcutaneous fat layer and stimulated dermal adipocyte regeneration in bleomycin-induced scleroderma skin. It reduced collagen deposition and dermal thickness, attenuating dermal fibrosis. Moreover, ad-organoids suppressed macrophage infiltration and promoted angiogenesis in the skin lesion. In conclusion, 3D culturing of MVFs with stepwise angiogenic and adipogenic induction is an effective strategy for the fabrication of ad-organoids, and the transplantation of prefabricated ad-organoids can improve skin sclerosis by restoring cutaneous fat and attenuating skin fibrosis. These findings offer a promising therapeutic approach for the treatment of localized scleroderma.

Combining High-Density Fat and Condensed Low-Density Fat Injections for Precise Facial Rejuvenation.

BACKGROUND: Facial aging involves ptosis, adipose atrophy, and skeletal resorption. Depletion of adipose tissue primarily affects the deep facial fat compartment, leading to facial depression or ptosis, accompanied by atrophy of the superficial compartment. Restoring volume in the deep fat compartment is crucial for facial rejuvenation, while enhancing its supportive properties is also important. The superficial fat compartment contains small-sized adipocytes, and autologous fat grafting is a popular approach. However, variability in fat retention, homogeneity, and processing methods can impact outcomes, necessitating careful selection of a suitable fat processing material for precise facial fat grafting. METHOD: A retrospective study was conducted on 50 patients who underwent facial augmentation using combined transplantation of high-density fat (HDF) and condensed low-density fat (CLDF) and 25 patients who underwent conventional Coleman fat grafting. Coleman fat was harvested by standard technique and the adipose tissue was divided into HDF and CLDF fractions through centrifugation. Subsequently, the low-density fat fraction was subjected to a process involving physical disruption followed by additional centrifugation to obtain CLDF. The CLDF fraction was consequently injected into the pre-SMAS subcutaneous layer of the superficial fat compartments. Patient satisfaction was evaluated using a typical Likert scale. Photographs were taken and imageological examinations were performed before and after treatment. RESULT: The CLDF+HDF grafting group demonstrated a significantly shorter duration of swelling (6.0 ± 1.2 to 12.6 ± 3.3 days) and higher level of patient satisfaction when compared to the Coleman fat group. No serious complications were observed among all the patients who received the injections. CONCLUSION: The use of this new treatment approach allows for precise fat transplantation in facial regions. The use of high-concentration fat filling for deep facial layers and CLDF filling for superficial layers is a safe and effective treatment plan for facial rejuvenation. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Type IV Collagen Promotes Adipogenic Differentiation of Adipose Stem Cells.

Type IV collagen is a major component of the extracellular matrix in adipose tissue. It is secreted during the lipogenic differentiation of mesenchymal stem cells, but its direct impact and mechanism on the differentiation of adipose-derived stem cells (ASCs) into lipids are unclear. In this study, ASCs were obtained from human liposuction samples and cultured. Lipogenic induction of ASCs was achieved using lipogenic induction medium. Immunofluorescence analysis revealed differential expression of type IV collagen during the early and late stages of adipogenic induction, displaying a distinct morphological encapsulation of ASCs. Silencing of type IV collagen using siRNA resulted in a significant decrease in adipogenic capacity, as indicated by reduced lipid droplet formation and downregulation of adipogenic-related gene transcription. Conversely, supplementation of the culture medium with synthetic type IV collagen demonstrated enhanced adipogenic induction efficiency, accompanied by upregulation of YAP/TAZ protein expression and its downstream target gene transcription. Furthermore, inhibition of the YAP/TAZ pathway using the inhibitor Blebbistatin attenuated the functionality of type IV collagen, leading to decreased lipid droplet formation and downregulation of adipocyte maturation-related gene expression. These findings highlight the crucial role of type IV collagen in promoting adipogenic differentiation of ASCs and suggest its involvement in the YAP/TAZ-mediated Hippo pathway.No Level Assigned This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Skeletal muscle provides a pro-browning microenvironment for transplanted brown adipose tissue to maintain its effect to ameliorate obesity in ob/ob mice.

Brown adipose tissue (BAT) transplantation is a promising means of increasing whole-body energy metabolism to ameliorate obesity. However, the changes in BAT following transplantation and the effects of the microenvironment of the recipient site on graft function have yet to be fully characterized. Therefore, we aimed to determine the effects of transplanting BAT from C57BL/6 mice into the dorsal subcutaneous region or deep to the quadriceps femoris muscle of leptin-deficient ob/ob mice. Subcutaneously transplanted BAT lost features of BAT and demonstrated greater inflammatory cell infiltration and more oil cysts 16 weeks following transplantation. By contrast, the sub-muscularly transplanted BAT maintained features of BAT and was more highly vascularized. Interestingly, sub-muscular BAT transplantation led to a significant increase in oxygen consumption and less inflammation in subcutaneous fat, which was associated with long-term reductions in insulin resistance and body mass gain, whereas the subcutaneous transplants failed after 16 weeks. These results demonstrate that the beneficial effects of BAT transplantation depend upon the microenvironment of the recipient site. Skeletal muscle may provide a microenvironment that maintains the inherent features of BAT grafts over a long period of time, which facilitates a reduction in obesity and improvements in glucose homeostasis.

Condensing of Low-Density Fat by Mechanical Process Improves Fat Retention and Reduces Oil Cyst Formation in Breast Reconstruction.

BACKGROUND: Although autologous fat grafting is a useful adjunct for breast reconstruction, its indications remain limited as large-volume fat grafting results in high absorption and complication rates. Low-density fat includes small numbers of viable cells and considerable oil, resulting in nodules and oil cysts. This study evaluated the volumization effect and complications with combined fat grafting of condense low-density fat and high-density fat. METHODS: This retrospective, single-center study included 25 patients who underwent combined grafting of condensed low-density fat and high-density fat (CLDF + HDF) and 20 patients who underwent conventional Coleman fat grafting for breast reconstruction from December 2017 to January 2022. Retention rates and complications were evaluated by magnetic resonance imaging and ultrasound rates. Patient satisfaction was evaluated using a typical Likert scale. Photographs were taken and imageological examinations were performed before and after treatment. OUTCOMES: Graft retention rate was higher in patients who underwent CLDF + HDF than Coleman fat grafting for breast reconstruction (38.40 ± 4.41% vs. 31.43 ± 5.43%, p <0.05). One patient in the CLDF + HDF grafting group, compared with twelve in the Coleman fat grafting group, developed oil cysts exceeding 1 cm. Patient satisfaction rate was higher in the CLDF + HDF grafting group. CONCLUSIONS: Mechanical processes can concentrate the cellular content of LDF and remove oil, condensing LDF to the level of HDF. Combined grafting of CLDF optimized by mechanical processing and HDF is effective for breast reconstruction, with a higher retention rate and a lower incidence of complications than conventional Coleman fat grafting. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Sequential Grafting of Fresh and Cryopreserved Fat After Mechanical Processing is a Safe and Effective Facial Rejuvenation Strategy.

BACKGROUND: The survival rate of fat transplants is variable and consequently multiple operations are often required to achieve satisfactory results. Fat cryopreservation technology is a good solution to this problem. At present, cryopreservation of fats needs to be added with cryopreservation agents, which brings unsafety and operational complexity to clinical applications. An efficient and safe strategy for fat cryopreservation must be developed. METHODS: A retrospective study was performed of all patients who underwent facial fat grafting and agreed to have their fat tissue cryopreserved from January 2018 to May 2021. Fat samples were physically processed to obtain SVF-gel, which was cryopreserved at - 20 °C for up to 3 months and injected after thawing. Images acquired by pre- and post-operative 3D scanning of the temporal region were compared to evaluate the retention rate of transplanted cryopreserved SVF-gel. RESULTS: No patients experienced serious complications after receiving cryopreserved fat transplantation. The retention rate of cryopreserved SVF-gel was 46.3 ± 7.7% at 3 months and 43.1 ± 7.2% at 6 months after transplantation. The swelling duration was significantly shorter after cryopreserved SVF-gel transplantation (5.5 ± 0.8 days) than after fresh fat transplantation (7.5 ± 0.7 days) (p < .031). CONCLUSION: Injection of cryopreserved SVF-gel achieves good retention rate for facial rejuvenation and has few side effects. Cryopreservation of SVF-gel is a safe and effective strategy for serial fat grafting for facial rejuvenation. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Use of brown adipose tissue transplantation and engineering as a thermogenic therapy in obesity and metabolic disease.

The induction of thermogenesis in brown adipose tissue is emerging as an attractive therapy for obesity and metabolic syndrome. However, the long-term efficacy and safety of clinical pharmaceutical agents have yet to be fully characterized. The transplantation of brown adipose tissue represents an alternative approach that might have a therapeutic effect by inducing a long-term increase in energy expenditure. However, limited tissue resources hinder the development of transplantation. Stem cell-based therapy and brown adipose tissue engineering, in addition to transplantation, represent alternative approaches that might resolve this problem. In this article, we discuss recent advances in understanding the mechanisms and applications of brown adipose tissue transplantation in the treatment of obesity and related metabolic disorders. Specifically, the induction of brown adipocytes and the fabrication of engineered brown adipose tissue as novel transplantation resources have long-term effects on ameliorating metabolic defects in rodent models. Additionally, we explore future prospects regarding the development of three-dimensional engineered brown adipose tissue and the associated challenges.

The browning and mobilization of subcutaneous white adipose tissue supports efficient skin repair.

Adipocytes in dermis are considered to be important participants in skin repair and regeneration, but the role of subcutaneous white adipose tissue (sWAT) in skin repair is poorly understood. Here, we revealed the dynamic changes of sWAT during wound healing process. Lineage-tracing mouse studies revealed that sWAT would enter into the large wound bed and participate in the formation of granulation tissue. Moreover, sWAT undergoes beiging after skin injury. Inhibition of sWAT beiging by genetically silencing PRDM16, a key regulator to beiging, hindered wound healing process. The transcriptomics results suggested that beige adipocytes in sWAT abundantly express neuregulin 4 (NRG4), which regulated macrophage polarization and the function of myofibroblasts. In diabetic wounds, the beiging of sWAT was significantly suppressed. Thus, adipocytes from sWAT regulate multiple aspects of repair and may be therapeutic for inflammatory diseases and defective wound healing associated with aging and diabetes.

Enhanced heterogeneous interface to construct intelligent conductive hydrogel gas sensor for individualized treatment of infected wounds.

In this study, we developed an enhanced heterogeneous interface intelligent conductive hydrogel NH3 sensor for individualized treatment of infected wounds. The sensor achieved monitoring, self-diagnosis, and adaptive gear adjustment functions. The PPY@PDA/PANI(3/6) sensor had a minimum NH3 detection concentration of 50 ppb and a response value of 2.94 %. It also had a theoretical detection limit of 49 ppt for infected wound gas. The sensor exhibited a fast response time of 23.2 s and a recovery time of 42.9 s. Tobramycin (TOB) was encapsulated in a self-healing QCS/OD hydrogel formed by quaternized chitosan (QCS) and oxidized dextran (OD), followed by the addition of polydopamine-coated polypyrrole nanowires (PPY@PDA) and polyaniline (PANI) to prepare electrically conductive drug-loaded PPY@PDA/PANI hydrogels. The drug-loaded PPY@PDA/PANI hydrogel was combined with a PANI/PVDF membrane to form an enhanced heterogeneous interfacial PPY@PDA/PANI/PVDF-based sensor, which could adaptively learn the individual wound ammonia response and adjust the speed of drug release from the PPY@PDA/PANI hydrogel with electrical stimulation. Drug release and animal studies demonstrated the efficacy of the PPY@PDA/PANI hydrogel in inhibiting infection and accelerating wound healing. In conclusion, the gas-sensitive conductive hydrogel sensing system is expected to enable intelligent drug delivery and provide personalized treatment for complex wound management.

Corrigendum: Spontaneous browning of white adipose tissue improves angiogenesis and reduces macrophage infiltration after fat grafting in mice.

[This corrects the article DOI: 10.3389/fcell.2022.845158.].

Spontaneous Browning of White Adipose Tissue Improves Angiogenesis and Reduces Macrophage Infiltration After Fat Grafting in Mice.

Background: Fat grafting is a frequently used technique; however, its survival/ regeneration mechanism is not fully understood. The browning of white adipocytes, a process initiated in response to external stimuli, is the conversion of white to beige adipocytes. The physiologic significance of the browning of adipocytes following transplantation is unclear. Methods: C57BL/6 mice received 150 mg grafts of inguinal adipose tissue, and then the transplanted fat was harvested and analyzed at different time points to assess the browning process. To verify the role of browning of adipocytes in fat grafting, the recipient mice were allocated to three groups, which were administered CL316243 or SR59230A to stimulate or suppress browning, respectively, or a control group after transplantation. Results: Browning of the grafts was present in the center of each as early as 7 days post-transplantation. The number of beige cells peaked at day 14 and then decreased gradually until they were almost absent at day 90. The activation of browning resulted in superior angiogenesis, higher expression of the pro-angiogenic molecules vascular endothelial growth factor A (VEGF-A) and fibroblast growth factor 21 (FGF21), fewer macrophages, and ultimately better graft survival (Upregulation, 59.17% ± 6.64% vs. Control, 40.33% ± 4.03%, *p < 0.05), whereas the inhibition of browning led to poor angiogenesis, lower expression of VEGF-A, increased inflammatory macrophages, and poor transplant retention at week 10 (Downregulation, 20.67% ± 3.69% vs. Control, 40.33% ± 4.03%, *p < 0.05). Conclusion: The browning of WAT following transplantation improves the survival of fat grafts by the promotion of angiogenesis and reducing macrophage.

Adipose-Derived Stem Cell-Enriched Lipotransfer Reverses Skin Sclerosis by Suppressing Dermal Inflammation.

BACKGROUND: Scleroderma is a chronic autoimmune disease with an incidence of 2.7 per 100,000 people. Traditional lipotransfer has been used to treat atrophic sclerotic skin. Enzymatically processed cell-assisted lipotransfer and mechanically processed stromal vascular fraction gel are fat products with abundant adipose-derived stem cells. This study assessed whether adipose-derived stem cell-enriched lipotransfer elicits superior therapeutic effects on scleroderma. METHODS: Scleroderma was induced in nude mice by injections of bleomycin for 4 weeks. Human-derived Coleman fat, cell-assisted lipotransfer, or stromal vascular fraction gel (0.1 ml) was injected into sclerotic lesions. Histologic examinations, terminal deoxynucleotidyl transferase dUTP nick end labeling, and expression analyses of inflammatory factors in skin lesions and transferred fat were performed at 4 weeks after implantation. RESULTS: Dermal thickness was lower in the groups injected with Coleman fat (339.0 ± 19.66 µm), cell-assisted lipotransfer (271.0 ± 16.15 µm), and stromal vascular fraction gel (197.8 ± 12.99 µm) than in the group injected with phosphate-buffered saline (493.3 ± 28.13 µm) ( p < 0.05). The numbers of terminal deoxynucleotidyl transferase dUTP nick end labeling + and Mac2 + cells in fat tissue were significantly higher in the group injected with Coleman fat than in those injected with stromal vascular fraction gel and cell-assisted lipotransfer. Expression of monocyte chemotactic protein-1 and interleukin-6 was significantly lower in the adipose-derived stem cell-enriched groups than in the Coleman fat group. Histologic analysis showed there were far fewer macrophages and myofibroblasts in skin lesions in the adipose-derived stem cell-enriched groups than in the Coleman fat group. CONCLUSIONS: Transplantation of stromal vascular fraction gel and cell-assisted lipotransfer, which contain abundant adipose-derived stem cells, reduces the levels of apoptotic cells and inflammation, significantly reverses skin sclerosis, and elicits superior anti-inflammatory and antifibrotic effects on scleroderma. CLINICAL RELEVANCE STATEMENT: This study provided an alternative adipose-based therapy, adipose-derived stem cell-enriched fat, for sclerotic lesions and showed its validity for interfering with the inflammation and fibrosis.

Corrigendum: Spontaneous browning of white adipose tissue improves angiogenesis and reduces macrophage infiltration after fat grafting in mice.

[This corrects the article DOI: 10.3389/fcell.2022.845158.].

A Preliminary Exploratory Study of Autologous Fat Transplantation in Breast Augmentation With Different Fat Transplantation Planes.

Background: Autologous fat transfer is common in breast augmentationor reconstruction. However, AFG recipient site in the breast for fat grafting has not been carefully investigated. Methods: Forty female patients requiring breast augmentation with fat grafting were randomly assigned into two groups. The retromammary group received 2/3 fat into the retromammary space and the other 1/3 into the subcutaneous and retropectoral planes. The retropectoral group received 2/3 fat into the retropectoral plane and the other 1/3 into the subcutaneous and retromammary planes. The fat grafting result at 6 months was assessed by 3D laser surface scanning and then ultrasound. Any complications were recorded during follow-up. Samples from a patient who underwent fat grafting for 6 months was obtained and histological examination was conducted. Results: No significant difference in the retention rate after 6 months was observed between the two groups (retromammary group: 35.9% ± 6.6; retropectoral group: 39.3% ± 5.1, p = 0.1076). The retromammary grouphad a higher incidence of oil cyst formation than the retropectoral group. Histological examination showed that there were more oil cysts and mac2 positive macrophage infiltration in the fat cells in retromammary group, while retropectoral group had more small-size adipocytes. Conclusion: Although fat grafting into the retropectoral plane did not provide a superior fat graft retention rate, it did lower the incidence of complications. The retropectoral space show great potential to become a favorable recipient site.

Improving Low-Density Fat by Condensing Cellular and Collagen Content through a Mechanical Process: Basic Research and Clinical Applications.

BACKGROUND: Large-volume fat grafting results in high absorption and complication rates. Low-density fat includes small numbers of viable cells and considerable oil, resulting in nodules and oil cysts. This study evaluated a strategy for large-volume fat grafting using a mechanical process to condense low-density fat and transplanting it with high-density fat. METHODS: Low-density fat, defined as the upper half of centrifuged lipoaspirates, was emulsified by intersyringe shifting and centrifuged to obtain condensed low-density fat. Fresh condensed low-density fat was analyzed by counting cells in the stromal vascular fraction, and by electron scanning and Western blotting. The retention rate and histologic changes of the product were analyzed using a fat grafting model in nude mice. Transplantation with a combination of condensed low-density fat and high-density fat was tested in patients undergoing breast reconstruction and breast augmentation. RESULTS: The condensed low-density fat derived from low-density fat contained a large number of stromal vascular fraction cells and collagens, comparable to that of high-density fat and much higher than in low-density fat and Coleman fat. Retention rates 12 weeks after transplantation were higher for condensed low-density fat (55.0 ± 7.5 percent) than for low-density fat (31.1 ± 5.7 percent) and Coleman fat (41.1 ± 6.8 percent), with condensed low-density fat having fewer oil cysts and lower macrophage infiltration. Patients grafted with combined condensed low-density fat and high-density fat showed good long-term volume retention. CONCLUSIONS: Using mechanical methods to condense low-density fat to a level comparable to that of high-density fat is a practical method of improving fat graft retention and avoiding severe complications. This new strategy may improve the quality of lipoaspirates for patients requiring large-volume augmentation.

Fat Grafting for Facial Rejuvenation Using Stromal Vascular Fraction Gel Injection.

This study used stromal vascular fraction gel (SVF-gel), a mechanically processed fat-derived product, to treat eye bag and tear trough deformity. SVF-gel is prepared by a process of centrifugation and intersyringe shifting and is particularly rich in SVF cells and native adipose extracellular matrix. SVF-gel injection is used alone or combined with transconjunctival eye bag removal. High satisfaction was noted among patients treated with SVF-gel injection for periorbital rejuvenation with fairly low complication rates. SVF-gel injection is a good alternative to assist transconjunctival lower eyelid blepharoplasty and correct the palpebromalar groove, tear trough deformity, and supraorbital hollow.

TLE3 represses colorectal cancer proliferation by inhibiting MAPK and AKT signaling pathways.

BACKGROUND: Transducin-like enhancer of Split3 (TLE3) serves as a transcriptional corepressor during cell differentiation and shows multiple roles in different kinds of cancers. Recently, TLE3 together with many other genes involved in Wnt/ß-catenin pathway were detected hyper-methylated in colorectal cancer (CRC). However, the potential role and the underlying mechanism of TLE3 in CRC progression remain scarce. METHODS: Gene expression profiles were analyzed in The Cancer Genome Atlas (TCGA) microarray dataset of 41 normal colorectal intestine tissues and 465 CRC tissues. Western blot and Real-time Quantitative PCR (RT-qPCR) were respectively performed to detect protein and mRNA expression in 8 pairs of CRC tissue and matched adjacent normal mucosa. Immunohistochemistry (IHC) was conducted to evaluate TLE3 protein expression in 105 paraffin-embedded, archived human CRC tissues from patients, whose survival data were analyzed with Kaplan-Meier method. In vitro experiments including MTT assay, colony formation assay, and soft agar formation assay were used to investigate the effects of TLE3 on CRC cell growth and proliferation. Additionally, subcutaneous tumorigenesis assay was performed in nude mice to confirm the effects of TLE3 in vivo. Furthermore, gene set enrichment analysis (GSEA) was run to explore potential mechanism of TLE3 in CRC, and then we measured the distribution of CRC cell cycle phases and apoptosis by flow cytometry, as well as the impacts of TLE3 on MAPK and AKT signaling pathways by Western blot and RT-qPCR. RESULTS: TLE3 was significantly down-regulated in 465 CRC tissues compared with 41 normal tissues. Both protein and mRNA expressions of TLE3 were down-regulated in CRC compared with matched adjacent normal mucosa. Lower expression of TLE3 was significantly associated with poorer survival of patients with CRC. Besides, knock down of TLE3 promoted CRC cell growth and proliferation, while overexpression of TLE3 showed suppressive effects. Furthermore, overexpression of TLE3 caused G1-S phase transition arrest, inhibition of MAPK and AKT pathways, and up-regulation of p21Cip1/WAF1 and p27Kip1. CONCLUSION: This study indicated that TLE3 repressed CRC proliferation partly through inhibition of MAPK and AKT signaling pathways, suggesting the possibility of TLE3 as a biomarker for CRC prognosis.