Institute Of Cancer Research Research Articles

Adherence to Cancer Prevention Lifestyle Recommendations Before, During, and 2 Years After Treatment for High-risk Breast Cancer

The American Institute for Cancer Research and American Cancer Society regularly publish modifiable lifestyle recommendations for cancer prevention. Whether these recommendations have an impact on high-risk breast cancer survival remains unknown. To investigate whether adherence to cancer prevention recommendations before, during, and 1 and 2 years after breast cancer treatment was associated with disease recurrence or mortality. The Diet, Exercise, Lifestyles, and Cancer Prognosis Study (DELCaP) was a prospective, observational cohort study designed to assess lifestyles before diagnosis, during treatment, and at 1 and 2 years after treatment completion, implemented ancillary to the Southwest Oncology Group (SWOG) S0221 trial, a multicenter trial that compared chemotherapy regimens in breast cancer. Participants were chemotherapy-naive patients with pathologic stage I to III high-risk breast cancer, defined as node-positive disease with hormone receptor-negative tumors larger than 1 cm or any tumor larger than 2 cm. Patients with poor performance status and comorbidities were excluded from S0221. The study was conducted from January 1, 2005, to December 31, 2010; mean (SD) follow-up time for those not experiencing an event was 7.7 (2.1) years through December 31, 2018. The analyses reported herein were performed from March 2022 to January 2023. An aggregated lifestyle index score comprising data from 4 time points and 7 lifestyles, including (1) physical activity, (2) body mass index, (3) fruit and vegetable consumption, (4) red and processed meat intake, (5) sugar-sweetened beverage consumption, (6) alcohol consumption, and (7) smoking. Higher scores indicated healthier lifestyle. Disease recurrence and all-cause mortality. A total of 1340 women (mean [SD] age, 51.3 [9.9] years) completed the baseline questionnaire. Most patients were diagnosed with hormone-receptor positive breast cancer (873 [65.3%]) and completed some education beyond high school (954 [71.2%]). In time-dependent multivariable analyses, patients with highest vs lowest lifestyle index scores experienced a 37.0% reduction in disease recurrence (hazard ratio, 0.63; 95% CI, 0.48-0.82) and a 58.0% reduction in mortality (hazard ratio, 0.42; 95% CI, 0.30-0.59). In this observational study of patients with high-risk breast cancer, strongest collective adherence to cancer prevention lifestyle recommendations was associated with significant reductions in disease recurrence and mortality. Education and implementation strategies to help patients adhere to cancer prevention recommendations throughout the cancer care continuum may be warranted in breast cancer.

STAT3 operates as a rheostat regulating sensitivity of NK cells to inflammatory cytokines

Abstract NK cells are innate lymphocytes that respond to a variety of cytokines during viral infection. Several homeostatic and inflammatory cytokines bind to receptors that signal via STAT3. STAT3 can form homodimers or heterodimers. To understand the role of STAT3 homodimers and heterodimers, we made use of knockout models in which NK cells lack STAT3 or specific cytokine receptors. We found that NK cells deficient in STAT3 expand less than wildtype NK cells during mouse cytomegalovirus (MCMV) infection. IL-21R and IL-10R are the only receptors expressed on NK cells that signal through STAT3 homodimers. However, IL-21R or IL-10R-deficient NK cells showed no or little expansion differences, suggesting additional cytokine receptors may be signaling through STAT3. To evaluate whether STAT3 regulates the sensitivity of NK cells to cytokines that signal through heterodimerization partners STAT1 and STAT5, we performed cytokine titrations. We found that sensitivity towards IFN-a and IL-15 was reduced in STAT3-deficient NK cells. NK cells require inflammatory cytokines for antiviral expansion but proliferate poorly in hyper-inflammatory environments. Because STAT3 appears to regulate the sensitivity of NK cells to inflammatory cytokines, we hypothesized that STAT3-deficient NK cells would be shielded in settings of hyper-inflammation. Indeed, using high-dose MCMV infection, we observed that STAT3-deficient NK cells expanded better than WT NK cells. RNA-seq analysis further corroborated that in high-dose infection STAT3-deficient NK cells are nearly indistinguishable from WT NK cells during regular-dose infection. Thus, we believe STAT3 operates as a rheostat, modulating the sensitivity of NK cells to their inflammatory environment. Supported by the Cancer Research Institute, Donald J. Gogel Postdoctoral Fellowship, CRI393

Adaptive immunity to ancient retroelements controls the skin tissue threshold of activation

Abstract A symbiotic dialogue between the microbiota and host is essential to promote protective immune responses in the skin. Integrated retroviral elements, or retrotransposons (RTNs), comprise up to 45% of the human genome. We previously found that immune responses to the skin microbiota are dependent on the reactivation and keratinocyte-intrinsic sensing of RTNs. Here, we explored the possibility that RTNs may also represent a source of antigens recognized by the skin adaptive immune system. We first established a model of sterile injury, where topical application of a mild detergent reactivated RTNs, and recruited CD8 T cells to the skin in an RTN-dependent manner. RNAseq analysis of RTN expression revealed Langerhans cells (LCs) have the most abundant RTN expression amongst skin dendritic cells. Our work demonstrated that CD8 T cell responses to sterile injury were entirely dependent on LCs, supporting the idea that LCs could present RTN antigens to CD8 T cells. To identify RTN-specific T cells, we selected peptides that were uniquely expressed in LCs, but not in thymic epithelial cells which mediate negative selection. Several peptides promoted cytokine production from injury-elicited CD8 T cells, confirming the presence of RTN-reactive CD8 T cells in the skin. RNA-seq analysis suggests RTN-specific CD8 T cells may contribute to wound repair and immunoregulation. Thus, mild stress, such as that induced by mild detergent, is sufficient to promote adaptive immunity to defined antigens derived from RTNs. Our work also proposes that homeostatic immunity to ancient retroviruses promotes a defined class of T cells responses aimed at maintaining barrier integrity in response to daily stressors encountered by the skin. Cancer Research Institute, Irvington Postdoctoral Fellowship

Plasticity of clonal memory CD8 T cell differentiation

Abstract Memory CD8 T cells provide highly specific and long-lasting protection against previously experienced pathogens. This potent protection is mediated by hundreds to thousands of unique clones with distinct T cell receptors (TCRs). Higher T cell clonal diversity and recognition of a wider range of pathogen antigens are associated with improved host protection. Therefore, understanding how clonal responses are organized is important for improving T cell function in a range of diseases. To comprehensively test the range of clonal memory T cell differentiation capacity, we deeply profiled polyclonal mouse memory CD8 T cells from acute lymphocytic choriomeningitis virus (LCMV) infection before and after viral rechallenge by paired single cell RNA and TCR sequencing. We found that individual memory T cell clone expansion and differentiation at the peak of the rechallenge response is highly variable. Clones are spread across various patterns of unique phenotypes ranging from highly cytotoxic to stem-like memory states. To quantify clonal differentiation plasticity, we transferred memory T cells into multiple new congenic recipients. Clonal progeny in different hosts acquired highly similar differentiation states suggesting that emergent memory clone behavior is imprinted by events during the primary infection. Indeed, transcriptional state of clones prior to rechallenge was sufficient to predict clone behavior upon rechallenge. This work provides a platform for understanding the mechanisms by which memory T cells make decisions and future studies will aim to perturb clones to elicit specific functions during a memory response. Supported by grants from the Burroughs Wellcome Fund (Career Award for Medical Scientists [ATS]), Cancer Research Institute (Technology Impact Award, Lloyd J. Old STAR Award [ATS]), American Society of Hematology (ASH Scholar Award [ATS]), Parker Institute for Cancer Immunotherapy (ATS), Stanford School of Medicine (Propel Postdoctoral Scholarship [KHG]), National Institutes of Health (5T32AI007290-38 [CJR])

Constitutive knock-in of MnSODK68R causes dilated cardiomyopathy and senescence in mice

The mitochondrial superoxide dismutase (SOD2 or MnSOD) is an essential enzyme for managing oxidative stress, particularly by removing the highly reactive superoxide generated during the normal functioning of the electron transport chain. The activity of MnSOD is regulated post-translationally by acetylation, specifically at lysine 68. The deacetylated form of MnSOD is catalytically active, and it has been shown that the deacetylation mimic, MnSOD K68R in which lysine 68 has been replaced with arginine, has upregulated dismutase activity in MCF7 and T47D human breast cancer cells. Overexpression of MnSOD has been shown to increase lifespan and protect against oxidative stress in the aging heart. We investigated a MnSOD K68R knock-in mouse with ubiquitous expression of the mutant enzyme and expected similar effects as the MnSOD overexpressing mice. However, we have found that MnSOD K68R homozygous knock-in mice develop dilated cardiomyopathy and have significantly decreased ejection fraction as evidenced by echocardiography at a young age. These mice also exhibit lower blood pressure compared to wild-type mice. Strikingly, these mice exhibit higher body weight, with increased lean mass and food consumption. Mitochondria isolated from MnSOD K68R knock-in mouse hearts showed higher respiratory capacity than WT counterparts. Biochemical analysis revealed increased lipid peroxidation, oxidized peroxiredoxin and senescence markers, possibly indicative of a premature aging phenotype. We conclude that MnSOD constitutive function upregulation may produce a sustained increase in hydrogen peroxide levels that could cause senescence and lead to dilated cardiomyopathy. This research was funded by NCI grant R01CA152601 and, R01CA214025, and the Cancer Prevention and Research Institute of Texas grant number RR20012. JS is supported by T32 AG 021890. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Heterologous prime-boost viral vector cancer vaccines have protective effects against a syngeneic mouse model of glioblastoma

Abstract Glioblastomas are immunologically “cold” tumors with intense immunosuppressive myeloid cell- and sparse cytotoxic T-cell infiltration. Therefore, viral vector vaccines may be a promising approach to boost the induction of glioblastoma-targeted T cells. It has been previously shown that heterologous prime-boost vaccination with chimpanzee-derived adenovirus ChAdOx1 and modified vaccinia Ankara (MVA) vectors can induce a high magnitude of CD8 +T cells specific for cancer-associated antigens and have therapeutic effects against mouse models of cancer. Therefore, we aimed to evaluate whether using ChAdOx1 and MVA vaccines targeting model antigen P1A, a mouse equivalent to human tumor-associated Melanoma Antigen GenE (MAGE)-type antigens, could have protective effects against a P1A-transfected BGL-1 glioblastoma model. Mice were vaccinated with ChAdOx1/MVA-P1A vaccines, then challenged subcutaneously with P1A-expressing BGL-1 cells. Prophylactic vaccination with ChAdOx1/MVA-P1A significantly prolonged the survival of syngeneic mice subcutaneously challenged with P1A-expressing BGL-1 tumors. Furthermore, different vaccination schedules significantly impacted the magnitude of antigen-specific CD8 +T-cell responses and protective efficacy. Future studies will investigate the protective effects of vaccination against intracranial P1A-expressing BGl-1 tumors. Supported by grants from the Ludwig Institute for Cancer Research, University of Oxford; the NIH Intramural Program (ZIA BC011877), the NIH Oxford-Cambridge Scholars Program, and the Marshall Scholarship Program.

Role of walnut-derived urolithins in reducing colonic inflammation in a patient cohort elucidated by an integrated omics analysis.

Abstract Diet can directly affect colon health and cancer risk. For example, the polyphenolic ellagitannins (punicigalin) that are present in walnuts are converted by the microbiome to a panel of bioactive urolithin metabolites. In particular, urolithin A has been studied for its anti-inflammatory and anti-cancer properties. In this pilot clinical trial, we evaluate the effects of walnut consumption (2-oz per day) on urinary urolithins and blood serum inflammatory markers, as well as on the transcriptome of normal colon and polyp tissue in 38 healthy obese and non-obese subjects. Our in-depth computational analysis of metabolomics and proteomics data suggests that subjects with higher urolithin A formation show significantly lower levels of several key serological markers of inflammation, including C-Peptide, sICAM-1, sIL6-R, Ghrelin, TRAIL, sVEGFR2, MCP2, and Haptoglobin. Furthermore, RNA-Seq transcriptomic analysis of colon tissue taken from proximal and distal colon shows reduced inflammatory programs and activation of the KICSTOR complex in subjects with higher urolithin A formation. These studies indicate that higher urolithin A formation is linked to an anti-inflammatory response, warranting further studies to better understand the role of urolithins in the inflammatory process. Supported by grants from American Institute for Cancer Research, California Walnut Commission, and National Cancer Institute (R01 CA252045).

A novel population of CD103-expressing CD56 +regulatory ILCs suppresses intratumoural T cells and are associated with poor prognosis in patients with ovarian carcinoma

Abstract Regulatory innate lymphoid cells (ILCregs) are an emerging family of immunoregulatory cells that suppress innate and adaptive immunity in the context of diseases, including cancer. However, there is currently a lack of lineage-defining surface markers or transcription factors (TFs) that distinguishes ILCregs from other subsets of ILCs. Moreover, the functional role of ILCregs within the tumour microenvironment (TME) has not been well established. In this study, we uncovered a novel population of CD103-expressing CD56 +ILCregs within the TME that correlated with poor recurrent-free survival (RFS) in patients with epithelial ovarian carcinoma (EOC). CD103 +ILCregs expressed distinct surface markers and TFs that distinguishes them from canonical CD56 +natural killer (NK) cells. Moreover, intratumoural CD103 +ILCregs expressed markers (i.e., CD39, CD69, CD101, TIGIT) and transcriptomic profiles that are shared with Tregs, suggesting that they may have immunoregulatory properties. We found that ascites-derived TGF-β drives NK cell conversion into CD103 +ILCs that can suppress autologous CD4 +T cells in vitro. Furthermore, intratumoural CD103 +ILCregs suppressed autologous CD8 +T cells in vitro. Finally, we found that the abundance of CD103 +ILCregs in the TME were associated with reduced levels of both activated CD4 +T cells and CD8 +T cells. Overall, we have identified CD103 +ILCregs as distinct immunoregulatory cells that suppresses T cells within the TME of patients with EOC. Further untangling of the complex networks of immune regulation, including ILCregs, may give rise to novel therapeutic targets to enhance anti-tumour immunity and improve clinical response rates to immunotherapies. This work was supported by a CIHR foundation award (CIHR FDN #143220), a TRANSCAN CIHR grant (CIHR – TRN #184713), a TRI grant from the Ontario Institute for Cancer Research (TRI OICR GCS #108796), and the Terry Fox Research Institute (TFRI-iTNT #1060). This project was also generously funded in memory of Mr. JIM JoTak under the Canadian Cancer Society (CCSRI #706152).

The CD16 and CD32b Fc-gamma receptors regulate antibody-mediated responses in mouse natural killer cells

Abstract Natural killer (NK) cells are innate lymphocytes capable of mediating immune responses without prior sensitization. NK cells express Fc-gamma receptors (FcγR) that engage the Fc region of IgG. Studies investigating the role of FcγR on mouse NK cells have been limited due to lack specific reagents. In this study, we characterize the expression and biological consequences of activating mouse NK cells through their FcγR. We demonstrate that most NK cells express the activating CD16 receptor, and a subset of NK cells also express the inhibitory CD32b receptor. Critically, these FcγRs are functional on mouse NK cells and can modulate antibody-mediated responses. We also characterized mice with conditional knock-out alleles of Fcgr3 (CD16) or Fcgr2b (CD32b) in the NK and innate lymphoid cell (ILC) lineage. NK cells in these mice did not reveal any developmental defects and were responsive to cross-linking activating NK receptors, cytokine stimulation, and killing of YAC-1 targets. Importantly, CD16-deficient NK cells failed to induce antibody-directed cellular cytotoxicity (ADCC) of antibody-coated B cell lymphomas in in vitro assays. In addition, we demonstrate the important role of CD16 on NK cells using an in vivo model of cancer immunotherapy using anti-CD20 antibody treatment of B cell lymphomas. This project was funded by fellowships from Burroughs Wellcome Fund (BWF) and the Cancer Research Institute to O.A.A., and grants from the Parker Institute for Cancer Immunotherapy (PICI), and the NIH (AI068129 and AI146581) to L.L.L.

Cryptic MHC-E epitope from influenza elicits a potent cytolytic T cell response

Abstract Non-classically-restricted T cells have shown remarkable therapeutic activity in a variety of disease settings; most notably, CD8 T cells restricted to MHC-E (Qa-1 in mice; HLA-E in humans) have proven capable of eliminating a primate lentivirus infection. Here, we used mass spectrometry to identify a Qa-1-restricted 9-mer peptide, named M-SL9, that elicits a strong CD8 T cell response in mice infected with influenza A virus. The M-SL9 specificity accounted for roughly 5–10% of all lung CD8 T cells, approaching the frequency of classically restricted CD8 T cell responses to flu. A broad investigation of the M-SL9-specific T cells showed that they are generally similar in phenotype to classically restricted CD8 T cells, including strong co-expression of type 1-associated cytokines (polyfunctionality) and cytolytic markers; however, M-SL9-specific T cells were unique in that they were anatomically restricted to the lungs and were not present in blood and spleen like other specificities. Intriguingly, the M-SL9 epitope derives from a previously unknown translation product encoded by the +1 alternative reading frame of the matrix 1 (M1) protein-coding sequence. Presentation and immunogenicity of M-SL9 were completely dependent on ribosomal initiation at the second AUG codon of the matrix gene segment. Analysis of potential precursors suggests that M-SL9 is processed from a novel 16-mer peptide that is not required for viral replication in mice. These discoveries imply that unconventional epitopes may drive a large fraction of physiologically relevant T cell responses, and M-SL9 presents a highly tractable system in which to dissect the mechanisms governing MHC-E-restricted T cell responses and their protective effects. Supported by Cancer Research Institute Irvington Fellowship, Roy and Diana Vagelos Molecular Life Sciences Program and College Alumni Society Research Grant from the Univ. of Pennsylvania, and NIH grants R01 AI146101, R01 AI153064, R01 AI046709-18S1, R01 AI046709, R01 AI125524, and R21 AI153978.

A membrane-associated inhibitory axis of MHC-I presentation for cancer immune evasion

Abstract Although the success of immune-checkpoint blockade has revolutionized cancer treatment, a number of tumors do not respond or develop resistance, including acute myeloid leukemia (AML). A potential mode of resistance is immune evasion of T-cell immunity involving aberrant MHC-I antigen presentation (AP). To map such mechanisms of resistance we identified key AP regulators using specific peptide-MHC-I-guided CRISPR/Cas9 screens in AML. The top-ranked negative regulators were surface protein Sushi Domain Containing 6 (SUSD6), Transmembrane Protein 127 (TMEM127), and the E3 ubiquitin ligase WWP2. SUSD6 is abundantly-expressed in AML and multiple solid cancers, ablation of which profoundly enhanced AP and reduced tumor growth in CD8 +T cell-dependent manner. Mechanistically, SUSD6 forms a molecular complex with TMEM127 and MHC-I, recruiting WWP2 for MHC-I lysosomal degradation. Together with the SUSD6-TMEM127-WWP2 gene signature negatively-correlated with cancer survival, our findings define a membrane-associated AP inhibitory axis as broadly-applicable therapeutic targets for both leukemia and solid cancers. This work is supported by the Mark Foundation ASPIRE Award to J.W. I.A. is supported by the NIH (NCI and NHLBI) (R01CA216421, R01 CA173636, 1R01CA228135, R01CA242020, 1R01HL159175), the Vogelstein Foundation and the Evans MDS Foundation. J.W. is supported by the NIH (R37CA273333-01, R21AI163924-02). Q.L. is supported by the Cancer Research Institute Irvington Postdoctoral Fellowship.

Effects of herbal tea (Platostoma palustre) on blood glucose regulation in vivo

Platostoma palustre jelly is a traditional food. Platostoma palustre has been used as folk medicine and is effective against heat-shock, hypertension and diabetes. Therefore, the aim of in vivo study was to determine the effects of herbal tea (Platostoma palustre) on blood glucose regulation. The commercial herbal tea (Platostoma palustre) was kindly provided by Yueta Agricultural Biotechnology Inc. Adult male 18 Institute of Cancer Research (ICR) mice [8 weeks old; body weight (BW) between 31-33 g] with specific pathogen-free conditions were used for this study. All ICR mice were divided respectively the normal control group (n = 6), the negative control group (n = 6), and the herbal tea group (n = 6). The ICR mice (the negative control group and the herbal tea group) were intraperitoneally injected with streptozotocin (65 mg/kg BW) and nicotinamide (230 mg/kg BW) for inducing the symptoms of hyperglycemia. In the herbal tea group, the herbal tea (10 mL/kg BW) was administrated to ICR mice by gavage. To monitor the blood glucose levels in ICR mice, blood was obtained from the tail of ICR mice, and blood glucose levels were determined using the external glucometer. Blood glucose measurements were conducted once in ICR mice before the experiment and were detected once per month after the experiment. Additionally, the change of ICR mice’ BW, the blood glucose of ICR mice, and the observation of ICR mice’ behavior were monitored and detected during the experiment. The results of this experiment showed · Weight change: the weight gain of the negative control group and the herbal tea group were significantly lower than those of the normal control group and there was no significant difference between the negative control group and the herbal tea group. · Fasting blood glucose and postprandial blood glucose: the fasting blood glucose of the negative control group was significantly higher than that of the normal control group, while the herbal tea group had a tendency to lower the fasting blood glucose, but there was no significant difference compared with the negative control group. The postprandial blood glucose level results showed that the negative control group was significantly higher than the normal control group. The postprandial blood glucose level of the herbal tea group was between the normal control group and the negative control group at the 4th week administration with the herbal tea. At the 8th week administration with the herbal tea, there was a significant difference compared with the negative control group. · Oral glucose tolerance test (OGTT): 4 to 12 weeks after hyperglycemia was induced, the blood glucose levels of the negative control group at each OGTT test point were significantly higher than those of the normal control group. The blood glucose level at the test point tended to decrease, but there was no significant difference from the negative control group. By calculating the area under the OGTT curve, it was found that the area under the curve of the negative control group was significantly higher than that of the normal control group, while the results of the herbal tea group at the 8th and 12th weeks were significantly lower than those of the negative control group. Taken all results together, the hyperglycemia was successfully induced in the experimental ICR mice. After administrating with the herbal tea, the fasting blood glucose level of the ICR mice tended to improve. In addition, a significant improvement was seen in the results of postprandial blood sugar and oral glucose tolerance test effectiveness. Therefore, based on the results of this experiment, it is speculated that drinking the herbal tea for 3 months has considerable potential for blood glucose regulation, which can be used as the basis for the development of related products of the herbal tea in the future.

Pharmacokinetics and bioavailability of curdione in mice by UPLC-MS/MS

AbstractA UPLC-MS/MS method was developed to determinate curdione in the mouse blood, and the pharmacokinetics of curdione in mice after intravenous (5 mg kg−1) and oral (20 mg kg−1) administration were studied. The HSS T3 column was used for separation, and column temperature was set at 40 °C. Multiple reaction monitoring (MRM) mode were used for determination of curdione. Blood samples were taken from the caudal vein of Institute of Cancer Research (ICR) mice after administration of curdione. It showed a good linear relationship in the range of 1–500 ng mL−1 (r > 0.998); the intra-day precision was <13%, the inter-day precision was <15%, and the accuracy was 90%–105%, the recovery was >77%, and the matrix effect was 97%–107%. The half-life was relatively short, and the bioavailability was 6.5%. The developed method was suitable for the pharmacokinetics of curdione in mice.

Prevention of Hepatocellular Carcinoma (HCC). White Paper of the Texas Collaborative Center for Hepatocellular Cancer (TeCH) Multi-stakeholder Conference

Texas has the highest age-adjusted incidence rate of hepatocellular carcinoma (HCC) in the United States. The Cancer Prevention and Research Institute of Texas has funded the Texas Collaborative Center for Hepatocellular Cancer (TeCH) to facilitate HCC research, education, and advocacy activities with the overall goal of reducing HCC mortality in Texas through coordination, collaboration, and advocacy. On September 17, 2022, TeCH co-sponsored a multi-stakeholder conference on HCC with the Baker Institute Center for Health and Biosciences. This conference was attended by HCC researchers, policy makers, payers, members from pharmaceutical industry and patient advocacy groups in and outside of Texas. This report summarizes the results of the conference. The goal of this meeting was to identify different strategies for preventing HCC and evaluate their readiness for implementation. We call for a statewide (1) viral hepatitis elimination program; (2) program to increase nonalcoholic steatohepatitis and obesity awareness; (3) research program to develop health care models that integrate alcohol associated liver disease treatment and treatment for alcohol use disorder; and (4) demonstration projects to evaluate the effectiveness of identifying and linking patient with advanced fibrosis and cirrhosis to clinical care.

Insights from establishing a high throughput viral diagnostic laboratory for SARS-CoV-2 RT-PCR testing facility: challenges and experiences.

Background: The World Health Organization declared the coronavirus disease 2019 (COVID-19) a global pandemic on 11 March 2020. Identifying the infected people and isolating them was the only measure that was available to control the viral spread, as there were no standardized treatment interventions available. Various public health measures, including vaccination, have been implemented to control the spread of the virus worldwide. India, being a densely populated country, required laboratories in different zones of the country with the capacity to test a large number of samples and report test results at the earliest. The Indian Council of Medical Research (ICMR) took the lead role in developing policies, generating advisories, formulating guidelines, and establishing and approving testing centers for COVID-19 testing. With advisories of ICMR, the National Institute of Cancer Prevention and Research (NICPR) established a high-throughput viral diagnostic laboratory (HTVDL) for RT-PCR-based diagnosis of SARS-CoV-2 in April 2020. HTVDL was established during the first lockdown to serve the nation in developing and adopting rapid testing procedures and to expand the testing capacity using "Real-Time PCR." The HTVDL provided its testing support to the national capital territory of Delhi and western Uttar Pradesh, with a testing capacity of 6000 tests per day. The experience of establishing a high-throughput laboratory with all standard operating procedures against varied challenges in a developing country such as India is explained in the current manuscript which will be useful globally to enhance the knowledge on establishing an HTVDL in pandemic or non-pandemic times.

MemU-Net: A new volumetric dose prediction model using deep learning techniques in radiation treatment planning

Objective:Automatic radiation treatment planning systems are gaining significance nowadays. This paper presents a novel deep learning architecture for automatically generating the radiotherapy dose information given to the cancer patients. Methods:A new deep learning architecture MemU-Net is proposed. MemU-Net incorporates the principles of two existing deep learning architectures: Memnet, which was proposed for image restoration, and U-Net, which was proposed for biomedical image segmentation. The proposed model is first trained and tested with the public dataset (OpenKBP). The predicted dose distributions are evaluated based on dosimetric values and Dose volume histogram measures. The results are compared with the predictions made by a standard 3D U-Net, Residual U-Net, Dense U-Net and 3D GAN. The statistical significance of the proposed model is tested by computing the paired t-test P-values. Finally the proposed model is trained and tested on a private dataset collected from MVR Cancer Center and Research Institute. The model performance was evaluated by plotting DVHs and analyzing dosimetric values. Results:The dose distributions of MemU-Net shows superiority in performance compared to other models. MemU-Net had a lower mean absolute error of the predicted and ground truth doses than other models for all targets and OARs except spinal cord. The dosimetric measures of the MemU-Net outperformed the state of the art models except in the case of D0.1cc of OARs. Conclusion:A new variant of U-Net, MemU-Net is proposed, that performs better than the state of the art models, in the dose prediction domain in radiotherapy.

Comparison the Difference of Thyroid Function Status between 2D & 3D CRT Treatment Technique in HNC Patients

Introduction: Thyroid gland is an important endocrine gland which regulates metabolic activities in our body as well as regulates vital body functions including breathing, heart rate, central and peripheral nervous systems, body weight, menstrual cycle, cholesterol levels and much more. As thyroid gland placed in the same field area treated by radiation therapy of non-thyroid head and neck cancer patient, so it is a challenge to preserve the normal thyroid function after completion of treatment. Objective: To assess the comparison the difference of thyroid function status between 2D & 3D CRT treatment technique in HNC patients. Methods: It was a quasi-experimental study was conducted at National Institute of Cancer Research & Hospital (NICRH), Mohakhali, Dhaka, Bangladesh from November 2016 and ended in October’2017. Total of 60 patients with above mentioned criteria were selected and each arm got 30 patients. Patient with histopathology report proven carcinoma of head and neck receiving radiotherapy were selected from tumor board of NICRH. The present study were carried out on patients with biopsy proven malignancies of head and neck who attended the Radiotherapy Department for radiotherapy and who met the selection criteria of the study. Results: Total study population was 60 among which 30 were in arm A (2D) and 30 were in arm B (3D). Majorities of the patients in arm A were 41-50 years old (46.7%). The mean age of the arm A patients was 52.27 (±7.38) years and that of the arm B was 49.50 (± 13.55) years. Thirty patients of head & neck cancers were included in arm A of the study, i.e. 2D arm. Out of which 21 (70%) were male and rest 9 (30%) were female. Thirty patients of head & neck cancers were included in arm B of the study, i.e. 3D arm. Out of which 23 (76.7%) were male and rest 7 (23.3%) were female. Most of the patients in both arms were farmers (40% in arm A and 60% in arm B). There were 9(30%) oropharyngeal, 8(26.7%) hypopharyngeal and 13(43.3%) ........

Aspirin microcrystals deposited on high-density microneedle tips for the preparation of soluble polymer microneedles.

To reduce mucosal damage in the gastrointestinal tract caused by aspirin, aspirin microcrystals were loaded in soluble polymeric microneedle (MN) tips. Aspirin was prepared into aspirin microcrystals by jet milling. Aspirin microcrystals with particle sizes of 0.5-5μm were loaded on MN tips with a height of 250µm or 300µm. The aspirin microcrystals suspended in a polymer solution were concentrated in the MN tips under negative pressure. The aspirin microcrystals had high stability in the MNs since they were not dissolved in solution during the fabrication process. The MN patch packaged in an aluminum-plastic bag containing silica gel desiccant can be stored at 4°C. The MN tips implanted in the skin of Institute of Cancer Research (ICR) mice dissolved within 30min. Isolated porcine ear skin was punctured by MNs with heights of 300μm and 250μm to depths of 130μm and 90μm, respectively. The fluorescent red (FR) release from MNs reached 98.59% within 24h. The MNs delivered aspirin microcrystals to the epidermis and dermis, providing a smooth plasma concentration in rats. The MNs loaded with aspirin microcrystals did not evoke primary irritation on the dorsal skin of Japanese white rabbits. In summary, MNs loaded with aspirin microcrystals provide a new approach to improve the stability of aspirin in MN patches.

Adherence to the World Cancer Research Fund/American Institute for Cancer Research cancer prevention recommendations throughout the life course and risk of colorectal cancer precursors.

Despite the increasing incidence in colorectal cancer (CRC) among the young population, the involvement of modifiable early-life exposures is understudied. We prospectively investigated the association of lifestyle score, which measures adherence to the 2018 World Cancer Research Fund/American Institute for Cancer Research (WCRF/AICR) cancer prevention recommendations, in adolescence and adulthood with risk of CRC precursors in 34,509 women enrolled in the Nurses' Health Study II. Participants reported adolescent diet in 1998 and subsequently underwent at least one lower gastrointestinal endoscopy between 1999 and 2015. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using multivariable logistic regression for clustered data. During follow-up (1998-2015), 3036 women had at least one adenoma, and 2660 had at least one serrated lesion. In multivariable analysis, per unit increase in adolescent WCRF/AICR lifestyle score was not associated with risk of total adenoma or serrated lesions, in contrast to adult WCRF/AICR lifestyle score (OR = 0.92, 95% CI: 0.87-0.97, Ptrend = 0.002 for total adenoma; and OR = 0.86, 95% CI: 0.81-0.92, Ptrend < 0.001 for total serrated lesions). Adherence to the 2018 WCRF/AICR recommendations during adulthood but not during adolescence was associated with a lower risk of CRC precursors.

Abstract 6733: Targeting Sp1 in Ewing sarcoma: a multi-approach method for the utilization of mithramycin

Abstract Purpose: Ewing Sarcoma (ES) is a bone and soft tissue cancer affecting young adults and children. Localized ES presents with a 5-year survival rate of 70%, while metastatic cases range from 15-30%. Our laboratory investigates combination treatments using less toxic agents to induce sensitization to chemotherapy in ES. The anti-cancer activity of an antineoplastic antibiotic, Mithramycin (Mit), against ES cells has been shown. Mit inhibits Specificity protein 1 (Sp1) a marker associated with aggressive cancer cell growth and resistance to chemo/radiation therapies. However, its mechanistic effects on other oncogenic proteins have yet to be elucidated in ES. The purpose of this study is to evaluate the effectiveness of Mit and combination with other chemotherapeutic agents, Etoposide and Vincristine, to inhibit ES cell growth and assess the effect on cancer related proteins regulated by Sp1. Future studies will expand upon Mithramycin's mechanism of action in Ewing Sarcoma utilizing RNA sequencing and computational methods. Methods: Anti-proliferative activity of Mit and/or Vincristine and Etoposide against ES cell lines, TC205 and CHLA10, was evaluated using CellTiterGlo kit. Dose curves and IC50 values were determined by Sigma-Plot software. The expression of Sp1 and survivin was determined by Western blot analysis. Cell lines were obtained from Children’s Oncology Group. The specific type of drug effect of the combination treatments were determined by analyzing the combination index obtained via Calcusyn software. Nude mice were injected with TC-205 cells and treated over two weeks with either Mit (1mg/kg per week) and/or Etoposide (5mg/kg per week) and tumor volume was compared. Protein models were obtained from RCSB PDB and homology tests were performed in the Swiss-model workspace. Results: Mit significantly decreased ES cell line viability and tumor volume in nude mice. Mit showed the ability to reduce the expression of Sp1 and offered differing effects on survivin expression, indicative of anti-apoptotic mechanisms being implemented in the ES cell lines. IC50 values of both chemotherapeutics and Mit were decreased by nearly 50% when used in combination and this effect was mirrored in further decreases in Sp1 expression. Synergistic drug responses were shown in the combination of Mit with both Vincristine and Etoposide. Sp1 and survivin protein models were established and homology verification using Ramachandran plots and QMEAN Z-scores indicated quality protein models for further computational studies. Conclusions: Mit may effectively sensitize ES cells and improve the response of chemotherapy while lowering necessary effective dosages. Studies to understand the mechanism of action of Mit on Sp1, survivin, and other proteins involved in Ewing Sarcomagenesis are underway. This study is partially supported by a grant from the Cancer Prevention and Research Institute of Texas (Award#: RP210046) Citation Format: Christoffer Lambring, Riyaz Basha, Umesh Sankpal. Targeting Sp1 in Ewing sarcoma: a multi-approach method for the utilization of mithramycin [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 6733.