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Using a panel of cancer cell lines, we characterized a novel degrader of AKT, MS21. In mutant PI3K–PTEN pathway cell lines, AKT degradation was superior to AKT kinase inhibition for reducing cell growth and sustaining lower signaling over many days. AKT degradation, but not kinase inhibition, profoundly lowered Aurora kinase B (AURKB) protein, which is known to be essential for cell division, and induced G2–M arrest and hyperploidy. PI3K activated AKT phosphorylation of AURKB on threonine 73, which protected it from proteasome degradation. A mutant of AURKB (T73E) that mimics phosphorylation and blocks degradation rescued cells from growth inhibition. Degrader-resistant lines were associated with low AKT phosphorylation, wild-type PI3K/PTEN status, and mutation of KRAS/BRAF. Pan-cancer analysis identified that 19% of cases have PI3K–PTEN pathway mutation without RAS pathway mutation, suggesting that these patients with cancer could benefit from AKT degrader therapy that leads to loss of AURKB.Significance:MS21 depletes cells of phosphorylated AKT (pAKT) and a newly identified AKT substrate, AURKB, to inhibit tumor growth in mice. MS21 is superior to prior agents that target PI3K and AKT due to its ability to selectively target active, pAKT and sustain repression of signaling to deplete AURKB.This article is highlighted in the In This Issue feature, p. 2945

Supplementary Data from Loss of PBRM1 Alters Promoter Histone Modifications and Activates ALDH1A1 to Drive Renal Cell Carcinoma

Supplementary Figure from AKT Degradation Selectively Inhibits the Growth of PI3K/PTEN Pathway–Mutant Cancers with Wild-Type KRAS and BRAF by Destabilizing Aurora Kinase B

Supplementary Table from AKT Degradation Selectively Inhibits the Growth of PI3K/PTEN Pathway–Mutant Cancers with Wild-Type KRAS and BRAF by Destabilizing Aurora Kinase B

Supplementary Figure from Loss of PBRM1 Alters Promoter Histone Modifications and Activates ALDH1A1 to Drive Renal Cell Carcinoma

Supplementary Data from AKT Degradation Selectively Inhibits the Growth of PI3K/PTEN Pathway–Mutant Cancers with Wild-Type KRAS and BRAF by Destabilizing Aurora Kinase B

Figure S1: Expression of GSH synthesis enzymes in ccRCC tumors and adjacent healthy kidney tissue, demonstrating increased GGT1 levels in ccRCC. Figure S2: GGT1 mRNA and protein abundance in ccRCC and renal epithelial cell lines. Figure S3: GGT1 inhibition results in proliferation defect in ccRCC cells. Figure S4: GGT1 knockdown results in cell cycle arrest in ccRCC cell lines. Figure S5: GSH levels and GSH/GSSG ratios are reduced in GGT1 KD tumor xenografts, relative to controls. Figure S6: GSH pathway inhibition leads to proliferation defects in ccRCC cells.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer. While the localized form of this disease can be treated surgically, advanced and metastatic stages are resistant to chemotherapies. Although more innovative treatments, such as targeted or immune-based therapies, exist, the need for new therapeutic options remains. ccRCC presents unique metabolic signatures and multiple studies have reported a significant increase in levels of reduced glutathione (GSH) and its precursors in ccRCC tumor samples compared with normal kidney tissues. These observations led us to investigate the effects of blocking the GSH pathway, particularly the gamma-glutamyltransferase 1 (GGT1) enzyme, in multiple ccRCC cell lines. In this study, we provide in vitro and in vivo evidence that GGT1/GSH pathway inhibition impacts ccRCC cell growth, through increased cell-cycle arrest. Of note, GGT1 inhibition also impairs ccRCC cell migration. Finally, pharmacologic GSH pathway inhibition decreases ccRCC cell proliferation and increases sensitivity to standard chemotherapy. Our results suggest that GGT1/GSH pathway inhibition represents a new strategy to overcome ccRCC chemoresistance.Implications:GGT1/GSH pathway inhibition represents a promising therapeutic strategy to overcome chemoresistance and inhibit progression of ccRCC tumors.

Using a panel of cancer cell lines, we characterized a novel degrader of AKT, MS21. In mutant PI3K–PTEN pathway cell lines, AKT degradation was superior to AKT kinase inhibition for reducing cell growth and sustaining lower signaling over many days. AKT degradation, but not kinase inhibition, profoundly lowered Aurora kinase B (AURKB) protein, which is known to be essential for cell division, and induced G2–M arrest and hyperploidy. PI3K activated AKT phosphorylation of AURKB on threonine 73, which protected it from proteasome degradation. A mutant of AURKB (T73E) that mimics phosphorylation and blocks degradation rescued cells from growth inhibition. Degrader-resistant lines were associated with low AKT phosphorylation, wild-type PI3K/PTEN status, and mutation of KRAS/BRAF. Pan-cancer analysis identified that 19% of cases have PI3K–PTEN pathway mutation without RAS pathway mutation, suggesting that these patients with cancer could benefit from AKT degrader therapy that leads to loss of AURKB. Significance: MS21 depletes cells of phosphorylated AKT (pAKT) and a newly identified AKT substrate, AURKB, to inhibit tumor growth in mice. MS21 is superior to prior agents that target PI3K and AKT due to its ability to selectively target active, pAKT and sustain repression of signaling to deplete AURKB. This article is highlighted in the In This Issue feature, p. 2945

The study was conducted to evaluate the clinical outcomes of various glaucoma surgeries (trabeculectomy, trabeculectomy with mitomycin-C and glaucoma valve implantation) in cases of refractory glaucoma. When it becomes obvious that medication will not help then alternatives should be tried.These alternatives can be in form of :- Filtration Surgery Termed as trabeculectomy allows drainage of aqueous humor from within the eye to underneath the conjunctiva where it is absorbed. Trabeculectomy with mitomycin C-The use of antifibrotic agents such as mitomycin-C and 5-fluorouracil, along with releasable sutures or laser suture lysis, enhances the longevity of guarded procedures, Drainage Implants Glaucoma drainage devices are designed to divert aqueous humor from the anterior chamber to an external reservoir, where a fibrous capsule forms about 4-6 weeks after surgery and regulates flow. Aims of Study 1. Study the clinical outcomes of trabeculectomy,trabeculetomy with mitomycin-C and glaucoma valve implantation in management of refractory glaucoma patients. 2.Evaluate the complications of glaucoma valve surgery. 3.Study the complications of trabeculectomy,trabeculectomy with mitomycin-C. The proposed study was conducted in upgraded department of ophthalmology,LLRM Medical College,Meerut. Population characteristics were number of eyes treated,diagnosed and follow up.The study was a prospective study with random selection of patients done from the OPD.The randomly selected patients were then divided into three groups, one group was subjected to trabeculectomy, second with trabeculectomy with mitomycin-C and the other group was subjected to glaucoma valve implantation. CRITERIA FOR FAILURE OF SURGERY: Ÿ IOP > 21 mm hg at end of 1 year or Ÿ IOP not reduced by 20% from baseline at end of 1 year or Ÿ IOP

Huge and reputed software industries are expected to deliver quality products. However, industry suffers from a loss of approximately $500 billion due to shoddy software quality. The quality of the product in terms of its accuracy, efficiency, and reliability can be revamped through testing by focusing attention on testing the product through effective test case generation and prioritization. The authors have proposed a test-case generation technique based on iterative listener genetic algorithm that generates test cases automatically. The proposed technique uses its adaptive nature and solves the issues like redundant test cases, inefficient test coverage percentage, high execution time, and increased computation complexity by maintaining the diversity of the population which will decrease the redundancy in test cases. The performance of the technique is compared with four existing test-case generation algorithms in terms of computational complexity, execution time, coverage, and it is observed that the proposed technique outperformed.

Tumors exhibit widespread enhancer landscape reprogramming compared to normal tissue. The etiology is believed to be largely cell-intrinsic in non-hormonal cancers, attributed to such genomic alterations as focal amplification of non-coding regions, aberrant activation of transcription factors, and non-coding mutations creating de novo transcription factor binding sites. Here, using freshly resected primary CRC tumors and patient-matched adjacent normal colon epithelia, we find divergent epigenetic landscapes between primary CRC tumors and CRC cell lines. We identify a unique super-enhancer signature largely absent in cell culture. Intriguingly, this phenomenon extends to highly recurrent aberrant super-enhancers gained in CRC over patient-matched normal epithelium suggesting novel insight into the etiology of enhancer reprogramming in CRC and its downstream relevance to tumor biology. We find one such super-enhancer activated in epithelial cancer cells due to surrounding inflammation in the tumor microenvironment. CRISPR-dcas9-KRAB interference of this super-enhancer identifies PDZK1IP1 as its target gene. PDZK1IP1 is previously observed to be highly up-regulated in CRC. However, the mechanism behind its transcriptional activation is not fully understood. We restore both the super-enhancer and PDZK1IP1 levels following treatment with cytokines or xenotransplantation into nude mice, thus demonstrating its etiology via local tumor microenvironment acquisition. Deletion of inflammatory transcription factors RELA and STAT3 in human CRC cells inhibits PDZK1IP1 induction in xenografts. PDZK1IP1 appears to be critical for CRC growth in the setting of its super-enhancer induction as xenografts, but not in cell culture where the super-enhancer is absent and expression is largely silent. Building on its known role in glucose uptake via SGLT receptors, we demonstrate mechanistically that PDZK1IP1 enhances the reductive capacity CRC cancer cells via the pentose phosphate pathway using polar metabolomic profiling. We show this activation enables efficient growth under oxidative conditions both in vitro and in vivo, challenging the previous notion that PDZK1IP1 acts as a tumor suppressor in CRC. Collectively, these observations highlight the biologic significance of epigenomic profiling on patient-matched primary specimens and identify this microenvironment-acquired super-enhancer as an oncogenic driver in the setting of the inflamed tumor. Citation Format: Royce Zhou, Jia Xu, Tiphaine Martin, Alexis Zachem, John He, Sait Ozturk, Deniz Demircioglu, Ankita Bansal, Andrew Trotta, Bruno Giotti, Berkley Gryder, Yao Shen, Saul Carcamo, Xuewei Wu, Kaitlyn Bosch, Benjamin Hopkins, Alexander Tsankov, Randolph Steinhagen, Drew Jones, John Asara, Jerry Chipuk, Rachel Brody, Steven Itzkowitz, Iok In Christine Chio, Dan Hasson, Emily Bernstein, Ramon Parsons. A local tumor microenvironment acquired super-enhancer induces an oncogenic driver in colorectal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3481.

Subunits of SWI/SNF chromatin remodeling complexes are frequently mutated in human malignancies. The PBAF complex is composed of multiple subunits, including the tumor-suppressor protein PBRM1 (BAF180), as well as ARID2 (BAF200), that are unique to this SWI/SNF complex. PBRM1 is mutated in various cancers, with a high mutation frequency in clear cell renal cell carcinoma (ccRCC). Here, we integrate RNA-seq, histone modification ChIP-seq, and ATAC-seq data to show that loss of PBRM1 results in de novo gains in H3K4me3 peaks throughout the epigenome, including activation of a retinoic acid biosynthesis and signaling gene signature. We show that one such target gene, ALDH1A1, which regulates a key step in retinoic acid biosynthesis, is consistently upregulated with PBRM1 loss in ccRCC cell lines and primary tumors, as well as non-malignant cells. We further find that ALDH1A1 increases the tumorigenic potential of ccRCC cells. Using biochemical methods, we show that ARID2 remains bound to other PBAF subunits after loss of PBRM1 and is essential for increased ALDH1A1 after loss of PBRM1, whereas other core SWI/SNF components are dispensable, including the ATPase subunit BRG1. In total, this study uses global epigenomic approaches to uncover novel mechanisms of PBRM1 tumor suppression in ccRCC. Implications: This study implicates the SWI/SNF subunit and tumor-suppressor PBRM1 in the regulation of promoter histone modifications and retinoic acid biosynthesis and signaling pathways in ccRCC and functionally validates one such target gene, the aldehyde dehydrogenase ALDH1A1.

Recently, blockchains have found a place in all sorts of industries, ranging from healthcare to the government sector, finance, real estate and utilities. Blockchain has proved to be advantageous to the applications which previously required a central authority and a trusted intermediary, as now blockchain allows them to operate in a decentralised manner and helps them to achieve the same functionality much more efficiently. Transactions in blockchains are trustless, that is, the involved parties without knowing each other and without establishing any trust relations, sit in any corner of the world and such transactions thus occur without any trusted middlemen. For authorisation of these transactions, strong cryptographic principles, which form an essential part of the blockchain infrastructure, are used. Smart contracts basically add a layer of logic and computation to the already existing trust infrastructure of the blockchain, and thus allows for appropriate, heavily automated and distributed workflows, to function efficiently. In this chapter, we outline the art of using smart contracts and blockchain technology to implement a decentralized, secure, transparent yet anonymous digital voting system.

Tumors exhibit widespread enhancer landscape reprogramming compared to normal tissue. The etiology is believed to be largely cell-intrinsic in non-hormonal cancers, attributed to such genomic alterations as focal amplification of non-coding regions, aberrant activation of transcription factors, and non-coding mutations creating de novo transcription factor binding sites. Here, using freshly resected primary CRC tumors and patient-matched adjacent normal colon epithelia, we find divergent epigenetic landscapes between primary CRC tumors and CRC cell lines. We identify a unique super-enhancer signature largely absent in cell culture. Intriguingly, this phenomenon extends to highly recurrent aberrant super-enhancers gained in CRC over patient-matched normal epithelium suggesting novel insight into the etiology of enhancer reprogramming in CRC and its downstream relevance to tumor biology. We find one such super-enhancer activated in epithelial cancer cells due to surrounding inflammation in the tumor microenvironment. We restore this super-enhancer and its expressed gene, PDZK1IP1, following treatment with cytokines or xenotransplantation into nude mice, thus demonstrating its etiology via local tumor microenvironment acquisition. Building on its known role in glucose uptake via SGLT receptors, we demonstrate mechanistically that PDZK1IP1 enhances the reductive capacity CRC cancer cells via the pentose phosphate pathway using polar metabolomic profiling. We show this activation enables efficient growth under oxidative conditions both in vitro and in vivo, challenging the previous notion that PDZK1IP1 acts as a tumor suppressor in CRC. Collectively, these observations highlight the biologic significance of epigenomic profiling on patient-matched primary specimens and identify this microenvironment-acquired super-enhancer as an oncogenic driver in the setting of the inflamed tumor.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer. While the localized form of this disease can be treated surgically, advanced and metastatic stages are resistant to chemotherapies. Although more innovative treatments, such as targeted or immune-based therapies, exist, the need for new therapeutic options remains. ccRCC presents unique metabolic signatures and multiple studies have reported a significant increase in levels of reduced glutathione (GSH) and its precursors in ccRCC tumor samples compared with normal kidney tissues. These observations led us to investigate the effects of blocking the GSH pathway, particularly the gamma-glutamyltransferase 1 (GGT1) enzyme, in multiple ccRCC cell lines. In this study, we provide in vitro and in vivo evidence that GGT1/GSH pathway inhibition impacts ccRCC cell growth, through increased cell-cycle arrest. Of note, GGT1 inhibition also impairs ccRCC cell migration. Finally, pharmacologic GSH pathway inhibition decreases ccRCC cell proliferation and increases sensitivity to standard chemotherapy. Our results suggest that GGT1/GSH pathway inhibition represents a new strategy to overcome ccRCC chemoresistance. Implications: GGT1/GSH pathway inhibition represents a promising therapeutic strategy to overcome chemoresistance and inhibit progression of ccRCC tumors.

Class Imbalance Problem is the major issue in machine intelligence producing biased classifiers that work well for the majority class but have a relatively poor performance for the minority class. To ensure the development of accurate prediction models, it is essential to deal with the class imbalance problem. In this paper, the class imbalance problem is handled using focused undersampling techniques viz. Cluster Based, Tomek Link and Condensed Nearest Neighbours which equalize the number of instances of the two types of classes by undersampling the majority class based on some particular criteria. This is in contrast to random undersampling where the data samples are selected randomly from the majority class leading to underfitting and loss of some important datapoints. To fairly compare and evaluate the performance of focused undersampling approaches, prediction models are constructed using popular machine learning classifiers like K-Nearest Neighbor, Decision Tree and Naive Bayes. The results have shown that Decision Tree outperformed other machine learning techniques. Comparing and contrasting the undersampling approaches for Decision Tree concluded Condensed Nearest Neighbours to be best amongst others.

Since the onset of COVID-19 pandemic, officials and several others have been making an effort to form informed decisions and take relevant measures to curb the outbreak. Mostly authorities have used standard statistical models, and epidemiological models to determine the outbreak. Although these models have shown to have accuracy in the past, they seem to be highly ineffective during the COVID-19 pandemic, mostly because of the complexities in it's outbreak. With the lack of studies done on the outbreak, it's imperative for us to determine the attributes which could show correlation with the spread for us to improve upon our predictions. Therefore in this paper the authors have tried to find the attributes which best help in the outbreak modelling and have applied those attributes to the traditional regression models to study their effects on the predictions.