•Salivary HPV DNA levels align with local disease burden in HPV oropharyngeal cancer.•The rise and fall of salivary HPV DNA often predicts treatment response or failure.•Plasma HPV cell-free DNA more ...often correlates with distant disease burden.
Quantifying tumor DNA in tissue and circulating in blood permits high-quality molecular monitoring to detect and track cancer progression. Evaluating tumor DNA in both blood and saliva in human papillomavirus (HPV)-associated oropharyngeal cancer (OPC) could provide a non-invasive and clinically actionable method for real-time disease detection.
We previously validated an ultrasensitive droplet-digital (dd)PCR assay targeting the dominant high-risk HPV subtypes causally linked to OPC. Here we enrolled an observational cohort to evaluate the predictive and prognostic potential of paired plasma-salivary tumor DNA among 21 patients with advanced HPV+OPC.
In patients with recurrent, persistent locoregional (LR) disease, median baseline normalized salivary HPV DNA was 10.9 copies/ng total DNA, nearly 20x higher compared with those with distant disease only (p = 0.01). A cutoff of 5 copies/ng yielded 87% sensitivity and 67% specificity for accurately predicting LR disease. Total tumor burden among those with LR disease strongly correlated with salivary HPV DNA levels (R = 0.83, p = 0.02). The rise and fall of salivary HPV DNA predicted treatment failure and response, respectively, in all patients with LR disease, and predated imaging findings. Among paired salivary-plasma (cell-free) cfDNA samples, only higher plasma HPV cfDNA levels were associated with poor outcomes (p < 0.01), suggesting that each bodily fluid provides unique information about HPV disease status.
Salivary HPV DNA provides valuable information about tumor burden and predicts treatment response in advanced HPV+OPC. Paired blood-saliva samples could be used to monitor HPV DNA with broad applications to inform diagnosis, prognosis, and surveillance in HPV-associated diseases.
Introduction
Improving surgical access in low- and middle-income countries is vital for the 5 billion people who lack safe surgical care. Tailoring a culturally sensitive approach to consent is ...essential for patient comprehension and comfort, thereby alleviating the effects of resource constraints and advancing equitable care. This study examines the consenting process for endoscopy at Kyabirwa Surgical Center in Kyabirwa, Jinja, Uganda, to assess patients’ knowledge and attitudes as a potential barrier to participating in endoscopic procedures.
Methods
All adult upper endoscopy (EGD) and colonoscopy patients were recruited to participate in a survey of their demographics, knowledge, and attitudes toward their procedure. All patients received a standard consultation explaining the procedure and its risks and benefits.
Results
75 patients were included; median age was 54 years and 56% (
n
= 42) were women. 92% (
n
= 69) of patients had never had an endoscopy before and 73% (
n
= 55) of patients were scheduled for an EGD while the remaining 27% (
n
= 20) were scheduled for a colonoscopy. Most patients 80% (
n
= 60) had a basic understanding of what an endoscopy is and 87% (
n
= 65) its diagnostic purpose. Few patients 15% (
n
= 11) knew of the most common side effects or if they would have a surgical scar 27% (
n
= 20). Overall, 46.7% (
n
= 35) of patients were moderately or severely fearful of getting an endoscopy. Additionally, 45.3% (
n
= 34) of patients were moderately or severely fearful of receiving anesthesia during their endoscopic procedure. Despite this fear, most patients 85.3% (
n
= 64) stated that they understood the benefits of the procedure either very well or extremely well.
Conclusions
Most patients understood the role that an endoscopic procedure plays in their care and its potential benefits. Despite this, many patients continued to have high levels of fear associated with both the endoscopic procedure and with receiving anesthesia during their procedure. Future patient education should focus on addressing patients’ fears and the risks of undergoing an endoscopy, which may improve the utilization of surgical services.
Acquired resistance to BH3 mimetic antagonists of BCL-2 and MCL-1 is an important clinical problem. Using acute myelogenous leukemia (AML) patient-derived xenograft (PDX) models of acquired ...resistance to BCL-2 (venetoclax) and MCL-1 (S63845) antagonists, we identify common principles of resistance and persistent vulnerabilities to overcome resistance. BH3 mimetic resistance is characterized by decreased mitochondrial apoptotic priming as measured by BH3 profiling, both in PDX models and human clinical samples, due to alterations in BCL-2 family proteins that vary among cases, but not to acquired mutations in leukemia genes. BCL-2 inhibition drives sequestered pro-apoptotic proteins to MCL-1 and vice versa, explaining why in vivo combinations of BCL-2 and MCL-1 antagonists are more effective when concurrent rather than sequential. Finally, drug-induced mitochondrial priming measured by dynamic BH3 profiling (DBP) identifies drugs that are persistently active in BH3 mimetic-resistant myeloblasts, including FLT-3 inhibitors and SMAC mimetics.
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•Reduced mitochondrial apoptotic priming drives acquired resistance to BH3 mimetics•BH3 profiling predicts clinical response to venetoclax and HMA combinations•Simultaneous BCL-2 and MCL-1 antagonism outperforms alternating regimens•Dynamic BH3 profiling identifies drug vulnerabilities in BH3 mimetic-resistant AML
Bhatt et al. demonstrate that resistance to BCL-2 and MCL-1 antagonists emerges via selection for reduced mitochondrial apoptotic priming. Rapid measurements of drug-induced apoptotic signaling measured by dynamic BH3 profiling identify targeted agents with in vivo efficacy. BCL-2 and MCL-1 antagonist combinations overcome resistance to either single agent.
Abstract
The development of targeted therapies has revolutionized the treatment of chronic lymphocytic leukemia (CLL). To date, these therapies are generally given continuously, indefinitely, leading ...to the development of resistance, which is often on target. Venetoclax is the first-in-class BCL-2 inhibitor which was initially approved for continuous therapy in relapsed high-risk CLL. In that context the BCL-2 G101V mutation (mut) was identified in post-progression samples and shown to reduce venetoclax binding to BCL-2, limiting its efficacy. The mut can be identified at low variant allele frequency (VAF) prior to clinical progression. We were therefore interested to identify the frequency of this mut in our cohort of relapsed refractory CLL patients (pts) on continuous venetoclax, and to assess the sensitivity of measurements in blood vs bone marrow. To this end we utilized a ddPCR assay which has LNA probes that specifically bind to either the BCL2 G101wt or G101V sequences, to screen for G101V muts in DNA extracted from patient samples. We also started to investigate additional co-occurring BCL2 muts in G101V positive samples by Sanger sequencing.
Our patient cohort included 28 pts, of whom 20 had serial samples collected during venetoclax therapy. The median age of the pts was 66, and they had a median of 3 prior therapies before venetoclax, including chemoimmunotherapy in 67.9% and a BTK inhibitor in 60.7%. Deletion of 17p was present in 43%, with five additional pts having isolated TP53 mut (total with known TP53 aberrancy, 61%). 75% (21/28) of those evaluated had an unmutated IGHV. The median duration of venetoclax treatment was 43.5 months (mos). The timing of the first sample tested was a median of 23.3 mos after venetoclax initiation.
We detected the G101V allele in peripheral blood mononuclear cells (PBMCs) in 9 out of 28 pts, at a median allele frequency (AF) of 1.38% (range 0.04%-22.31%), at a median of 44.6 mos on venetoclax. Out of the three pts who had G101V detected at multiple timepoints, two had AF increased with time (7.8 fold increase over 6 mos and 7.7 fold increase over 5 mos, respectively), one had similar AF with time (4.68% at 18.9 mos, 3.43% at 23.8 mos on treatment). Six of these pts also had bone marrow evaluated and all were also positive (at a median AF of 0.21%; range 0.2%-18.66%); one additional patient without a PBMC sample at that timepoint was positive in bone marrow. In order to screen for any co-occurring acquired resistance muts in BCL2 G101V positive samples, we performed Sanger sequencing across the BCL2 open-reading frame. We have identified a duplication mut (R107-R110dup) in one of the samples.
In conclusion, this study shows that approximately one-third of pts on continuous venetoclax for 2+ years develop evidence of low-level BCL-2 G101V mut. Further work is underway to identify additional co-existing muts in BCL2 or other genes, and to characterize the additional genetic events at the time of clear clinical progression.
Citation Format: Yanan Kuang, Stacey M. Fernandes, Rayan Fardoun, Kevin Vasquez, Abhishek Mogili, Cloud P. Paweletz, Jennifer R. Brown. BCL-2 G101V mutations develop in one-third of patients on continuous venetoclax abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3960.
The objective of this work is to incorporate a Look-up table based analytical model of TFET using Verilog-A in Cadence Design framework and thus using this integrated model we are able to design and ...simulate some of the digital circuits. The Look-up-tables are obtained from TCAD simulations. III-V materials are taken into consideration, as these devices are having higher driving capability as per ITRS Norms. A library is created in Cadence and based on N-type TFET P-type TFET, Inverter, NAND, NOR, half adder are realized. A comparative analysis is made with the state of arts of FinFET using 22nm technology. We proposed an Inverter circuit using hybrid TFET-FinFET. As the TFET technology can attain a low subthreshold slope of nearly 40mV/decade, we opted for this technology for designing. Therefore, this promotes the TFET-based circuits to work with a 0.5V supply voltage which leads to a reduction in dynamic power consumption and leakage current when compared to the current CMOS technology. As the results suggest that many more advantages and we can choose TFET for designing. The schematics and circuit simulation is done using the Cadence Virtuoso tool.
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