The Krüppel-like factors (KLFs) are a family of transcriptional regulators that play crucial roles in regulating diverse cellular processes including development, proliferation, differentiation, metabolism, and carcinogenesis across various tissues. KLFs play pivotal roles in gastrointestinal pathologies, and exhibit tissue- and cell-type-specific expression patterns throughout the gastrointestinal tract. During gastrointestinal (GI) development, KLFs orchestrate the transition from embryonic to adult gene programming, with specific family members being essential for proper organogenesis and tissue formation. KLFs also function as context-dependent modulators of GI homeostasis, inflammation, and carcinogenesis in adult tissues and interact with major signaling pathways such as PI3K/AKT, NF-κB, Wnt, Notch, MAPK, and TGF-β. This review comprehensively examines the roles of KLFs in GI health and disease, focusing on their expression patterns, regulatory mechanisms, function in normal homeostasis, and therapeutic implications for gastrointestinal disorders.

Recent developments in neural radiance field (NeRF) processing have leveraged the power of neural networks to quickly reconstruct 3D spaces from 2D images. Our objective was to utilize this technology to 3D render video recordings of diagnostic cystoscopies and test their fidelity. With institutional review board (IRB) approval, we recorded two diagnostic cystoscopies, one with an Ambu single-use flexible cystoscope and the other with a Richard Wolf digital cystoscope. We converted the videos to images and manually curated approximately 100 representative images, which minimized blur and spanned a large segment of the bladder. We then utilized the NVIDIA Instant Neural Graphics Primitives (iNGP), a NeRF algorithm that uses multiresolution hash encoding with a compact neural network for significantly faster convergence, to reconstruct the bladder and render novel, unseen views within seconds. We computed the structural similarity index (SSIM) and peak signal-to-noise ratio (PSNR) to assess the quality and fidelity of the 3D rendering. Both videos were able to be utilized for 3D rendering using iNGP. The rendering derived from the Richard Wolf cystoscopy had a PSNR = 29.8 (min = 27.2, max = 32.6) and SSIM = 0.89. Similarly, the rendering derived from the Ambu cystoscopy had a PSNR = 31.3 (min = 27.1, max = 35.1) and SSIM = 0.90. Independent of cystoscopy equipment, both 3D renderings achieved reasonable fidelity. Major limitations to widespread adoption of this technology include the need for a curator to select representative and high-quality images from the initial cystoscopy video recording and the relatively small segments of bladder successfully rendered. Nonetheless, we feel that with further refinement, this technology can be scaled to create 3D renderings of cystoscopies that will enable evaluation of both completeness and quality of the cystoscopy. Furthermore, this technology would be able to facilitate the comparison of cystoscopies performed in the same patient over time.

Type 1 diabetes (T1D) results from insulin insufficiency due to the loss or dysfunction of pancreatic beta cells following T cell-mediated autoimmune attack. Currently the only long-term therapy is daily exogenous insulin replacement. The ideal curative approach is the durable restoration of functional islets via transplantation. To date the limiting factors impeding realization of this goal is the lack of a cost effective and limitless source of high-quality islets suitable for transplantation and the ability to provide long-term islet graft acceptance without prolonged need for deleterious immunosuppression. Ongoing clinical trials are testing islets derived from human induced pluripotent stem cells (iPSC); however, long-term acceptance of islet graft will require a effective therapeutic strategy to prevent engrafted islet destruction by pre-existing islet-antigen specific T cells. Here we demonstrate in the NOD mouse model for T1D that autologous islet graft acceptance can be achieved by the targeted elimination of (re)-activated islet-reactive CD4 + and CD8 + T effector (Teff) cells in the initial post-transplantation period by using a short-acting, combination therapy that results in the elimination of islet-reactive Teff cells by exacerbation of their natural DNA damage response (DDR) to drive apoptosis while at the same time maintaining endogenous Treg cells. Article Highlights:Activated beta-cell reactive CD4 + and CD8 + T effector cells undergo a profound DNA-damage response which is targetable by small molecule inhibitors of the p53 and cell cycle pathways that lead to apoptosis. The use of a combination of MDM2 and WEE1 inhibitors, which termed "p53 potentiation with checkpoint abrogation" (PPCA), conferred significant therapeutic efficacy in treating mouse models of new onset T1D. Specific targeting of these T effector cells by PPCA results in a loss of inflammatory T cell subsets, notably proliferation CD4 + Th0 and Th1 subsets and CD8 + T effector memory cells, as determined by single cell RNA-seq studies with the preservation of T regulatory cells. When autologous islet grafts are given to established diabetic NOD mice, a single course of PPCA results in long-term islet graft acceptance, restoration of normoglycemia and loss of beta cell specific CD4 + and CD8 + T cells. PPCA shows promise as a potential means of estimating islet graft tolerance in T1D recipients of islet graft transplantation.

The transcriptional regulator Krüppel-like factor 5 (KLF5) is highly expressed in squamous epithelial cells of the esophagus. Increased KLF5 activity induces tumorigenesis and promotes metastasis in several cancers, although this function appears to be context-dependent. Here, we demonstrate that acute KLF5 inhibition, both genetically and with the potent KLF5 inhibitor ML264, causes non-transformed human primary esophageal squamous epithelial cells to enter the epithelial to mesenchymal transition (EMT). Moreover, chronic KLF5 inhibition with ML264 leads to the development of cells with a mesenchymal phenotype characterized by the expression of mesenchymal markers and functionally by reduced cell growth and increased migration and cellular invasion. This EMT resulting from chronic KLF5 inhibition is not driven by β-Catenin or TGF-β signaling. Pharmacologically, ML264 inhibits KLF5 by promoting proteasomal-mediated degradation. Taken together, we demonstrate that reduced KLF5 activity reprograms epithelial cells towards a mesenchymal phenotype and enhances their migratory and invasive potential. These findings have potential implications not only for esophageal cancers but also for normal processes such as esophageal tissue repair following injury.

Abstract Metastasis is a major contributor to cancer morbidity and mortality. However, studying this complex, multi-step, multi-organ process is challenging. There is a pressing need to dissect the contributions of the tumor microenvironment (TME) on this process in a controlled and precise manner. Fortunately, the utilization of microfluidic “organs-on-chips” (OOC) in cancer research is facilitating such investigations. Moreover, by combining organoids and organs-on-chips we can enhance the progress of these studies in a patient specific manner. In this study, we have developed a cancer-on-chip model aimed at investigating early metastatic spread within the colon milieu. To achieve this, we employ colorectal cancer (CRC) cell lines as well as patient-derived CRC organoids chosen from our biorepository to encompass diverse representations across race/ethnicity, sex, and mutational profiles. The tumor cells are introduced into the upper chamber of the OOC model, while human microvascular endothelial cells (HIMECs) are introduced into the lower chamber to establish a tube-like structure resembling a blood vessel. These two chambers are separated by a porous membrane, and the chip is flanked by vacuum chambers to introduce stretch-like motions, simulating peristalsis in the gastrointestinal track. Creating a mechanically dynamic microenvironment facilitates the exploration of neurotransmitters and their impact on tumor cell behavior. Recent studies have suggested that tumor cells may exploit physiological processes, such as neurotransmitter signaling, to their benefit. By utilizing mass spectrometry-based metabolomics we detected dynamic and patient organoid-specific changes in neurotransmitter levels (i.e., serotonin, aspartate, glutamate, γ-aminobutyric acid (GABA)) in the effluent of our CRC-on-chip model. Coupled with live-cell imaging, we discovered tumor cell-derived GABA, a major inhibitory neurotransmitter, serves as an energy source for supporting tumor cell intravasation. This finding was most pronounced in KRAS-mutant tumor cells and further supported by analysis of CRC patient samples from The Cancer Genome Atlas (TCGA) database. We were able to reverse the GABA-mediated invasion effect by inhibiting 4-aminobutyrate aminotransferase (ABAT), the enzyme responsible for GABA catabolism. In summary, our cancer-on-chip model holds promise for exploring various aspects of the metastatic process and uncovering potential therapeutic targets, such as neurotransmitters. Citation Format: Carly Strelez, Rachel Perez, John S. Chlystek, Christopher Cherry, Bethany Haliday, Ah Young Yoon, Curran Shah, Ren X. Sun, Roy Lau, Aaron Schatz, Josh Neman, Heinz Josef-Lenz, Jonathan Katz, Shannon M. Mumenthaler. Involvement of GABAergic signaling in enhanced tumor cell invasion in a mechanically dynamic tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1507.

You have accessJournal of UrologySurgical Technology & Simulation: Artificial Intelligence II (PD27)1 May 2024PD27-06 3D RENDERING OF CYSTOSCOPY VIDEO FOOTAGE: A NOVEL METHOD UTILIZING NEURAL RADIANCE FIELD PROCESSING Jonathan E. Katz, Jamie Finnegan, Jingpei Lu, Shan Lin, Michael Yip, and Roger Sur Jonathan E. KatzJonathan E. Katz , Jamie FinneganJamie Finnegan , Jingpei LuJingpei Lu , Shan LinShan Lin , Michael YipMichael Yip , and Roger SurRoger Sur View All Author Informationhttps://doi.org/10.1097/01.JU.0001008580.58088.27.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Recent developments in Neural Radiance Field Processing (NERF) have leveraged the power of neural networks to quickly reconstruct 3D spaces from 2D images. Our objective was to utilize this technology to 3D render video recordings of diagnostic cystoscopies and test their fidelity. METHODS: With IRB approval we recorded two diagnostic cystoscopies (one with an Ambu disposable flexible cystoscope and the other with a Wolf digital cystoscope). We converted the videos to images and then curated the images to choose approximately 100 images, which minimized blur and spanned a large segment of the bladder. We then utilized the NVIDIA Instant Neural Graphics Primitives (iNGP), a NeRF algorithm that uses multiresolution hash encoding with a compact neural network for significantly faster convergence, to reconstruct the bladder and render novel, unseen views. We computed the structural index similarity (SSIM) and Peak signal to noise ratio (PSNR) to assess the quality and fidelity of the 3D rendering. RESULTS: Both videos were able to be utilized for 3D rendering using iNGP. The rendering derived from the Wolf cystoscopy had a PSNR=29.8 [min=27.2, max=32.6] and SSIM=0.89. Similarly, the rendering derived from the Ambu cystoscopy had a PSNR=31.3 [min=27.1, max=35.1] and SSIM=0.90, static sample images below (Image 1). CONCLUSIONS: Independent of cystoscopy equipment, both 3D renderings achieved reasonable fidelity. Major limitations to widespread adoption of this technology include the need for a curator to select representative and high-quality images from the initial cystoscopy video recording and the relatively small segments of bladder successfully rendered. Nonetheless, we feel that with further refinement this technology can be scaled to create 3D renderings of cystoscopies that will enable evaluation of both completeness and quality of the cystoscopy. Furthermore, this technology would be able to facilitate the comparison of cystoscopies performed in the same patient over time. Download PPT Source of Funding: None © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e552 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Jonathan E. Katz More articles by this author Jamie Finnegan More articles by this author Jingpei Lu More articles by this author Shan Lin More articles by this author Michael Yip More articles by this author Roger Sur More articles by this author Expand All Advertisement PDF downloadLoading ...

Alloreactive memory T cells have been implicated as central drivers of transplant rejection. Perplexingly, innate cytokines, such as IL-6, IL-1β, and IL-12, are also associated with rejection of organ transplants. However, the pathways of innate immune activation in allogeneic transplantation are unclear. While the role of microbial and cell death products has been previously described, we identified alloreactive memory CD4 T cells as the primary triggers of innate inflammation. Memory CD4 T cells engaged MHC II-mismatched dendritic cells (DCs), leading to the production of innate inflammatory cytokines. This innate inflammation was independent of several pattern recognition receptors and was primarily driven by TNF superfamily ligands expressed by alloreactive memory CD4 T cells. Blocking of CD40L and TNFα resulted in dampened inflammation, and mice genetically deficient in these molecules exhibited prolonged survival of cardiac allografts. Furthermore, myeloid cell and CD8 T cell infiltration into cardiac transplants was compromised in both CD40L- and TNFα-deficient recipients. Strikingly, we found that priming of naive alloreactive CD8 T cells was dependent on licensing of DCs by memory CD4 T cells. This study unravels the key mechanisms by which alloreactive memory CD4 T cells contribute to destructive pathology and transplant rejection.

Affinity Purification of EGFR, JUN, JNK, and FOS

β ‐TrCP is an E3 ubiquitin ligase that plays important roles in multiple human cancers including esophageal squamous cell carcinoma (ESCC). Analysis of ESCC patient samples reveal that only protein level but not transcript level of β ‐TrCP associated with patient prognosis, suggesting regulators of β ‐TrCP protein stability play an essential role in ESCC progression and may be novel targets to develop ESCC therapies. Although β ‐TrCP stability is known to be mediated by the ubiquitin‐proteasome system, it is unclear which enzymes play a major role to determine β ‐TrCP stability in the context of ESCC. In this study, OTUD6B is identified as a potent deubiquitinase of β ‐TrCP that suppress ESCC progression through the OTUD6B‐ β ‐TrCP‐SNAIL axis. Low OTUD6B expression is associated with a poor prognosis of ESCC patients. Importantly, all‐trans retinoic acid (ATRA) is found to promote OTUD6B translation and thus suppress ESCC tumor growth and enhance the response of ESCC tumors to anti‐PD‐1 immunotherapies. These findings demonstrate that OTUD6B is a crucial deubiquitinase of β ‐TrCP in ESCC and suggest combination of ATRA and anti‐PD‐1 immune checkpoint inhibitor may benefit a cohort of ESCC patients.

Supplementary Figure 2 from Haploinsufficiency of Krüppel-Like Factor 4 Promotes Adenomatous Polyposis Coli–Dependent Intestinal Tumorigenesis

Supplementary Figure 1 from Haploinsufficiency of Krüppel-Like Factor 4 Promotes Adenomatous Polyposis Coli–Dependent Intestinal Tumorigenesis