Enhanced BDNF and ROS in Mucosa of Lower Motor Neuron Lesioned Dog Bladder Following Somatic Motor Nerve Transfer

AUTHORS

Nagat Frara, Kais Jawawdeh, Dania Giaddui, Istvan P. Tamas, Ryan P. Gares, Elizabeth R. McGonagle, Brendan A. Hilliard, Mikhail A. Kolpakov, Lewis Bright-Rowe, Alan S. Braverman, Justin M. Brown, Michael R. Ruggieri, Sr., Mary F. Barbe

ABSTRACT

Neurotrophic factors and reactive oxygen species (ROS) modulate neuronal plasticity. In a model of a lower motor neuron lesioned bladder, somatic nerve transfer was used as a reinnervation strategy. Levels of neurotrophins, ROS, and TNF-α in bladder mucosa and muscle layers collected from three groups of adult female dogs: (1) Decentralized, via bilateral transection of coccygeal and sacral spinal roots, lumbar 7 dorsal roots, and hypogastric nerves, then 6–21 mo recovery; (2) reinnervated (ObNT-Reinn), after similar decentralization for 12 mo, then bilateral obturator-to-vesical nerve transfer and 8–12 mo recovery; and (3) Controls. In mucosa, BDNF and ROS levels were highest in ObNT-Reinn bladders, GDNF and TNF-α levels were restored to Control levels in ObNT-Reinn bladders (lowest in Decentralized). NT-3 and ARTN were lower in ObNT-Reinn and Decentralized bladders versus Controls. In muscle, ROS was lower in ObNT-Reinn muscle versus Controls. BDNF mucosa levels correlated with bladder axonal density and detrusor layer thickness; and GDNF mucosal correlated with bladder contraction after vesical or transferred obturator nerve electrical stimulation, as did BDNF and GDNF muscle levels. The increased BDNF and GDNF in bladders that underwent somatic nerve transfer with subsequent recovery suggest that BDNF and GDNF may help promote the reestablishment of bladder innervation.

Podoplanin-positive cell-derived small extracellular vesicles contribute to cardiac amyloidosis after myocardial infarction

AUTHORS

Maria Cimini, Ulrich H.E. Hansmann, Carolina Gonzalez, Andrew D. Chesney, May M. Truongcao, Erhe Gao, Tao Wang, Rajika Roy, Elvira Forte, Vandana Mallaredy, Charan Thej, Ajit Magadum, Darukeshwara Joladarashi, Cindy Benedict, Water J. Koch, Çağla Tükel, Raj Kishore

ABSTRACT

Cardiac amyloidosis is a secondary phenomenon of an already pre-existing chronic condition. Whether cardiac amyloidosis represents one of the complications post myocardial infarction (MI) has yet to be fully understood. Here, we show that amyloidosis occurs after MI and that amyloid fibers are composed of macrophage-derived serum amyloid A 3 (SAA3) monomers. SAA3 overproduction in macrophages is triggered by exosomal communication from cardiac stromal cells (CSCs), which, in response to MI, activate the expression of a platelet aggregation-inducing type I transmembrane glycoprotein, Podoplanin (PDPN). CSCPDPN+-derived small extracellular vesicles (sEVs) are enriched in SAA3, and exosomal SAA3 engages with macrophage by Toll-like receptor 2, triggering overproduction with consequent impaired clearance and aggregation of SAA3 monomers into rigid fibers. SAA3 amyloid deposits reduce cardiac contractility and increase scar stiffness. Inhibition of SAA3 aggregation by retro-inverso D-peptide, specifically designed to bind SAA3 monomers, prevents the deposition of SAA3 amyloid fibrils and improves heart function post MI.

Roles of Necroptosis, Apoptosis, and Inflammation in Colorectal Carcinogenesis: A Longitudinal Human Study

AUTHORS

Timothy Su, Xiangzhu Zhu, Yong Li, Chang Yu, Xinqing Deng, Eugene Shubin, Lifang Hou, Jing Zhao, Lei Fan, Heping Zhang, Harvey J. Murff, Reid M. Ness, Martha J. Shrubsole, Qi Dai

ABSTRACT

Necroptosis triggers an inflammatory cascade associated with antimicrobial defense. No prospective human study has yet explored the role of necroptosis in colorectal cancer development. We conducted quantitative analysis of biomarkers for necroptosis [transient receptor potential cation channel subfamily M member 7 (TRPM7) and phosphorylated mixed lineage kinase domain-like protein], inflammation [cyclooxygenase-2 (COX-2)], apoptosis [BCL2-associated X (BAX) and terminal deoxynucleotidyl transferase dUTP nick end labeling], and cell proliferation (Ki67). This was done using tissue microarray biospecimens from the Cooperative Human Tissue Network and rectal biopsies from a longitudinal study within the Personalized Prevention of Colorectal Cancer Trial. In the human colorectal adenoma–carcinoma sequence, we observed an inverse expression trend between BAX and TRPM7; TRPM7 decreased from normal mucosa to small and large adenomas but significantly increased in early colorectal cancer stages (Ptrend = 0.004). It maintained high levels through all cancer stages. An increased COX-2 intensity in the epithelium was noted during tumorigenesis (Ptrend = 0.02) and was significantly associated with an elevated risk of metachronous polyps (odds ratio = 3.04; 95% confidence interval, 1.07–8.61; Ptrend = 0.02). The combined composite index scores of TRPM7 and COX-2 were strongly linked to 6- to 47-fold increased risks for metachronous adenoma/serrated polyps, whereas combined scores of phosphorylated mixed lineage kinase domain-like protein or TRPM7 with BAX were associated with an 11.5- or 13.3-fold elevated risk for metachronous serrated polyps. In conclusion, our findings suggest that COX-2 expression within normal-looking colorectal mucosa is significantly associated with an increased risk of metachronous colorectal polyp. Furthermore, our results propose the hypothesis that synergistic interactions among necroptosis, inflammation, and apoptosis could play a pivotal role in human colorectal tumorigenesis.

Prevention Relevance: Our findings suggest that COX-2 expression and combined scores of COX-2, TRPM7, and BAX hold promise for predicting the risk of metachronous polyps and could potentially serve as a tool for assessing the effectiveness of chemopreventive agents in preventing colorectal cancer during intervention trials.

Mesenchymal Stem Cells-Derived Extracellular Vesicles Mimetics as Osteoinductive Mediators for Bone Healing

AUTHORS

Antoine Karoichan, Ling Li, Celine J. Agnes, Bettina M. Willie, Maryam Tabrizian

ABSTRACT

Bone defects remain challenging to treat, with common therapies still relying on invasive approaches. Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) offer a promising alternative due to their regenerative and immunomodulatory properties, but challenges in scalable EV production limit clinical translation. Nanoghosts (NGs) are an emerging class of EV-mimetics synthesized through the physical distortion of ghost cells that offer innate bioactivity similar to EVs while having more scalable yields. In this study, the osteogenic potential of NGs made from MSC ghosts (MSC-NGs) is explored for the first time and contrasted with conventional MSC-EVs. MSC-NGs are generated through sonication, yielding two-fold more vesicles compared to MSC-EVs from the same number of cells. Unlike MSC-EVs, MSC-NGs significantly enhanced the osteogenic differentiation of MSCs, evidenced by increased alkaline phosphatase (ALP) activity and early mineralization. Proteomic analysis further revealed that MSC-NGs are more enriched in osteogenesis-related proteins than MSC-EVs. In vivo, treatment of a 0.5 mm mouse femoral osteotomy with MSC-NGs accelerated fracture healing, showing increased callus mineralization by day 14 and improved bone marrow reconstitution by day 21, along with reduced osteoclastic activity. These findings demonstrate MSC-NGs as scalable and effective therapeutics for bone tissue engineering, offering advantages over MSC-EVs in future bone healing strategies.

Arthroscopy and arthrotomy were equally effective for medial meniscal release but arthroscopy was...

Arthroscopy and arthrotomy were equally effective for medial meniscal release but arthroscopy was minimally advantageous at preventing iatrogenic damage to the caudal cruciate ligament: a canine cadaveric study

AUTHORS

Dana N. Gale, Steven W. Frederick, Bianca F. Hettlich, Jennifer J. Au, Tatiana Motta

ABSTRACT

Objective: To compare efficacy of four techniques used for medial meniscal release by medial caudal meniscotibial ligament transection and evaluate associated iatrogenic damage to the medial meniscus, caudal cruciate ligament (CdCL), and articular cartilage of the canine stifle joint.

Study design: Twenty-four pairs of canine cadaveric pelvic limbs were randomly assigned to groups by methods of approach, cranial tibial translation, and meniscal release. I: arthrotomy, Hohmann, #11 scalpel blade; II: arthrotomy, Hohmann, #64 Beaver blade; III: arthroscopy, Hohmann, meniscal hook knife; IV: arthroscopy, no joint translation, meniscal hook knife. Post-procedure stifle dissection and evaluation of meniscal release success rate and presence of iatrogenic damage were performed. Fisher’s exact tests were performed for meniscal release and damage comparisons. Iatrogenic damage to the CdCL and articular cartilage were compared using generalized linear mixed effects model and linear mixed effects models (G/LMM) respectively.

Results: Incomplete meniscal release was identified in 0/12 (0%) stifles in group I, 1/12 (8.3%) stifles in group II, 2/12 (16.7%) stifles in group III, and 1/12 (8.3%) stifles in group IV (p = 0.89, Fisher’s exact test). There was no difference in iatrogenic meniscal damage rates between groups (p = 0.48, Fisher’s exact test). There were no differences in total surface area of iatrogenic articular cartilage damage in any tested region between groups: femoral trochlea (p = 0.32, LMM), femoral condyles (p = 0.54, LMM), tibia (p = 0.28, LMM). Group I had more iatrogenic damage to the CdCL than group IV (p = 0.04, GLMM); no other differences were found.

Conclusion: Arthroscopy and arthrotomy were equally effective for performing medial meniscal release by transection of the medial caudal meniscotibial ligament. Arthroscopic evaluation and medial meniscal release without joint translation was minimally advantageous in preventing iatrogenic damage to the CdCL.

Yes-associated protein plays oncogenic roles in human sporadic colorectal adenomas

AUTHORS

Lei Fan, Xingyi Guo, Mary K Washington, Jiajun Shi, Reid M Ness, Qi Liu, Wanqing Wen, Shuya Huang, Xiao Liu, Qiuyin Cai, Wei Zheng, Robert J Coffey, Martha J Shrubsole, Timothy Su

ABSTRACT

The role of Hippo-Yes-associated protein (YAP) in human colorectal cancer (CRC) presents contradictory results. We examined the function of YAP in the early stages of CRC by quantitatively measuring the expression of phospho-YAPS127 (p-YAP) and five APC-related proteins in 145 sporadic adenomas from the Tennessee Colorectal Polyp Study, conducting APC sequencing for 114 adenomas, and analyzing YAP-correlated cancer pathways using gene expression data from 326 adenomas obtained from Gene Expression Omnibus. The p-YAP expression was significantly correlated with YAP expression (r = 0.53, P < .0001) and nuclear β-catenin (r = 0.26, P = .0018) in adenoma tissues. Both p-YAP and nuclear β-catenin were associated with APC mutations (P = .05). A strong association was observed between p-YAP overexpression and advanced adenoma odds (OR = 12.62, 95% CI = 4.57–34.86, P trend < .001), which persisted after adjusting for covariates and biomarkers (OR = 12.31, 95% CI = 3.78–40.10, P trend < .0001). P-YAP exhibited a sensitivity of 77.4% and specificity of 78.2% in defining advanced versus nonadvanced adenomas. Additionally, synergistic interaction was noted between p-YAP positivity and nuclear β-catenin on advanced adenomas (OR = 16.82, 95% CI = 4.41–64.08, P < .0001). YAP-correlated genes were significantly enriched in autophagy, unfolded protein response, and sirtuin pathways showing predominantly pro-tumorigenic alterations. Collectively, YAP plays an oncogenic role in interacting with Wnt as well as other cancer pathways within human sporadic adenomas. P-YAP could be a potential biomarker for human high-risk sporadic adenomas.