Soluble cytokine biomarkers are the master regulators of cell-to-cell communication-- the means by which one cell tells another what to do or where to go. Determining the biomarker signatures of immune cell populations reveals essential insights into cellular functions, for example, when probing new immuno-oncology targets or investigating treatment responses in patients. This webcast will illustrate how biopharma researchers are using the multiplex Luminex® platform in a high-throughput manner to accelerate immuno-oncology discovery research from target discovery through preclinical development. A focus will be on novel soluble checkpoint panels, the integration of biomarker data with other analytics platforms, and strategies to handle large multiplex datasets from dozens of different programs.

Learning Objectives:

1. Benefits of using multiplexed biomarker profiling from early to late stages of an immuno-oncology discovery project

2. Generating high quality discovery data using multiplex protein detection assays

3. How biomarker data helps solidify target biology and targeting rationale

While circulating cell-free DNA (ccfDNA) and to some extend CTCs from blood are routinely used as analyte in liquid biopsy cancer research applications, circulating cell-free RNA (ccfRNA) has recently also gained relevance for biomarker studies. The combination of insights from different analytes promises an increase in understanding molecular processes of tumor biology. However, there are still challenges to overcome in the preanalytical workflow which e.g. includes defining suitable blood collection tubes, transport concepts or extraction methods. In this presentation, we show the multimodal use of the PAXgene Blood ccfDNA Tube for plasma extraction as part of a wholistic liquid biopsy workflow.

Learning Objectives:

1. Workflow standardization in Liquid biopsy research

2. ccfDNA stabilization

3. Multmodality in liquid biopsy research 

Multiplex detection of oncogenic mutations using LNA-based assays on the QIAcuity digital PCR system Digital PCR (dPCR) enables specific and sensitive detection of genetic alterations in oncogenes. It is particularly suited for the analysis of rare mutations in precious samples such as circulating cell-free DNA (ccfDNA) from blood and other body fluids. For reliable detection of single base exchanges at low frequency, both, a reproducibly working dPCR system and optimized PCR assays, are indispensable. Locked Nucleic Acid (LNA)-enhanced oligonucleotides with substantially high levels of affinity for their complementary sequences make these assays excellent tools for the detection and discrimination of highly similar DNA targets. Here, we discuss simultaneous, high-sensitivity detection of up to three different mutations using QIAGEN’s QIAcuity dPCR system in combination with the dPCR LNA Mutation Assays.

Learning Objectives:

1. Learn how to detect simultaneous, high-sensitivity detection of up to three different mutations 

2. Learn how the QIAcuity DPCR System works

3. Learn what are dPCR LNA mutations assays

The continued rapid expansion of immunotherapies, including both in vivo and ex vivo therapeutics, has driven the development and adoption of novel tools to study, asses and understand these therapies. Not least among these new tools are cell lines that can be engineered to more faithfully represent the biological state of cells, both healthy and diseased, that these immunotherapies will come into contact with. Tumor associated antigens provide a substantial and specific set of targets for immunotherapies. However, extracting these cells from patient samples provides a sample which has a huge number of variables, making it difficult to determine when and why a specific immunotherapy might fail. Alternatively, immortalized cell lines overexpressing such antigens can provide an unrealistic assessment of therapeutic efficacy. Our team utilized it's advanced cell engineering workflow to generate cell lines which express controlled levels of tumor-associated antigens and introduced these expression systems into biologically-relevant cells. These modified cell lines can be used as controls in validated assays or for the development of new potency assays for testing candidate immunotherapies, and provide established levels of antigen expression that can help provide deeper insight into the function and performance of novel therapeutics.

Learning Objectives:

1. Understand how engineered, biologically relevant cell lines are useful to faithfully represent the state of cells immunotherapies will come into contact with

2. Understanding how controlled levels of antigen presenting panels can assess and advance Immunotherapies

Martin highlights various factors impacting molecular analysis from FFPE samples and key challenges and considerations while working with this precious yet challenging sample type.

Learning Objectives:

1. Nucleic acid preparation from FFPE samples

2. Key considerations to achieve and verify high yields and quality 

Antibodies have become one of the most important tools in life science research, allowing the detection, quantitation, and determination of changes in proteins and other molecules with respect to time and subcellular location.  MilliporeSigma ZooMAb® antibodies represent a new generation of monoclonal antibodies that are specifically engineered using proprietary technologies that provide state-of-the-art consistency and applications performance.  ZooMAb® antibodies are designed with the most user-friendly formulation, handling, and storage features available today and have been validated in multiple immunoassay applications.  This platform acquired its unique name because of its proprietary B cell immortalization technology that also allows us to produce monoclonal antibodies from a wide range of species including rabbits, goats, llama, sheep, and more.  The monoclonal antibodies produced and selected are subsequently converted into a eukaryotic recombinant expression format.  Thus, all ZooMAb® antibodies are offered as affinity purified recombinant proteins for highest reproducibility, purity, and performance.  In this webinar, we will highlight the development process of ZooMAb® antibodies, show examples of the outstanding affinity, avidity, and specificity of these antibodies in multiple immunoassays, provide details about the earth friendly format they are provided in, and finally deliver details about future directions as well as custom development opportunities.

Learning Objectives:

1. Learn about advantages of recombinant antibodies in cancer research

2. Learn about ZooMAb antibodies  - MilliporeSigma's recombinant monoclonal antibody line

Immunohistochemistry (IHC) is used to determine spatial relationships where we can identify the localization of target proteins in specific regions and cells of a tissue.  IHC has been the gold standard to analyze protein expression analysis in tissues; however, the IHC assay may not always work as not all antibodies are robust reagents for detection of target proteins.  For that reason, we have utilized several orthogonal assays to analyze localization of proteins and mRNA of targets within cells where robust antibody reagents are not available or specificity is in question. I will briefly describe several effective techniques that have helped my pathobiology team in the method development of assays that are used for target validation and characterization of human diseases and animal disease models.

Learning Objectives:

1. The presentation will provide the audience the many tools available for the method development of antibodies by utilizing orthogonal methods to ensure specificity of the IHC assay or equivalent histology-related assay

2. The presentation will help you discern how to resolve common problems encountered in immunohistochemical staining

Mutations in the epidermal growth factor receptor gene (EGFR) and the known Kirsten rat sarcoma viral oncogene  (KRAS) are significant drivers in the development of non-small-cell lung cancer (NSCLC). Treatment is typically guided by which mutations are present, and there are several possible mutations that can arise during treatment, so detecting them quickly and reliably is key to helping the patient. With NSCLC, biopsied patient samples tend to be very small, which can hamper many conventional genetic tests. Thus the ideal test in NSCLC is one that can work with minimal inputs, and test for several mutations at the same time. This webcast will present a new liquid biopsy test using multiplexing assays that contained mutant variants. The QX ONE Droplet Digital PCR (ddPCR) System allows for automated operation and advanced multiplexing to detect mutations in EGFR and KRAS using as little as 5 ng of cell-free DNA extracted from plasma.

Learning Objectives:

1. What are the key benefits to having a high-throughput, multiplexing ddPCR system?

2. Are the results the same between standard duplex reactions and multiplexed reactions?

3. What is the greatest number of samples that can be processed to results overnight on the QX200 vs the QX ONE?

Individuals of under-represented minority ancestry are at disproportional risk for higher incidence and mortality rates for particular cancers. The unequal burden of cancer in certain racial and ethnic groups, known as “cancer disparities”, can be attributed to a multilevel interplay among neighborhood and population-wide social, psychosocial, lifestyle, environmental, health system, and biological determinants of health. ARS is a key step in gene expression enabling individual genes to encode multiple proteins, and is emerging as a major driver of abnormal phenotypic heterogeneity including gaining aggressive characteristics and tolerance to anticancer therapy. RNA splicing–related genetic and genomic variation in tumors includes oncogenes dysregulated by ARS, spliceosome-dependent transformation, RNA splicing–related immunogenic epitopes and race–related cancer aggressiveness and drug response. Our laboratory has conducted a number of studies in multiple cancers stratified by race (both self-identified and by ancestral genotyping) using either deep RNAseq or array-based technologies on retro- and prospectively banked tissue samples, as well as computational analysis of publicly available databases such as TCGA. We have shown that the burden of race-related differential ARS (D-ARS) is much higher than differential gene expression, and that many of the D-ARS are functionally involved in oncogenic signaling pathways and statistically associate with survival. We have built an application, “CanSplice”, to mine genomic datasets for D-ARS by patient race and ethnicity. We have also begun to identify and test, preclinically and clinically, approaches to modulate and exploit ARS for therapeutic application, including splice-switching oligonucleotides, small molecules targeting RNA splicing or RNA splice variants, and combination regimens with immunotherapies. Our findings identify race-related ARS targets that may aid in the development of new biomarkers and precision therapeutic agents that have the potential to mitigate cancer disparities.

Learning Objectives:

1. Define the various contributors to cancer health disparities and some points of intersectionality between them

2. Explain the potential role of RNA splicing in the biology of ancestry-related difference in the molecular etiology of cancer in different populations

3. Identify approaches to pharmacologically target RNA splicing to mitigate cancer aggressiveness

Every patient is different and so is every tumor! The term “personalized oncology” today is largely associated with sequencing approaches. Current publications proof more and more that only few people really benefit from sequencing their tumor genome leaving many patients underrepresented and underserved. With Reverse Clincial Engineering, we have designed an organoid-based, multi-omics driven pipeline that supports us in understanding each tumor the best possible way in a clinical setting. Timing is crucial when it comes to treating cancer patients. It has been well established, that chances of cure dramatically decrease from treatment line to treatment line. Using organoids for drug screening has been shown to be feasible by many groups for various tumor entities. We use multiple regions of the donor tumor to generate distinct patient-derived 3D (PD3D®) organoids in less than 4 weeks, screening all drugs that are considered by the oncologist, plus drugs that may have an approval for different tumor entities. In parallel, we perform DigiWest, a bead-based multiplexed western blot based on Luminex’ xMAP technology platform. Here we can interrogate a set of up to 200 (phospho) protein specific antibodies in parallel. In contrast to RNAseq, this delivers not only information about the expression, but also of the activation of signaling pathways, with the benefit of a significantly less complicated infrastructure in comparison to sequencing. From all sources, we compile a patient-specific dataset including histo-pathological data, FISH analysis, sequencing data, drug screening data and proteomic data we compile a comprehensive report for each patient which is reported to the tumor board to be discussed within a multidisciplinary team. In conclusion: We rethink oncology and use complementory technologies to put the patient in the focus, enabling patient and oncologist to make informed decisions at the right time.

Learning Objectives:

1. Show that a “sequencing alone” approach is not in the patient’s best interest

2. Convince the audience that using a multi layered omics approach helps understanding the biology and as such opening new treatment options

Prostate cancer is the most common non-cutaneous cancer affecting men in the US and the second leading cause of cancer mortality. Prostate cancer is also growing as a source of cancer morbidity and mortality globally, with the greatest relative burden of lethal disease born by men in West and sub-Saharan Africa, and in the Caribbean. Age-standardize mortality rates in the US have fallen 50% in the era of PSA-based early detection, but African American men continue to face a roughly 2.5-fold excess burden of lethal disease compared to Caucasian men. The disparity is worst among younger African American men, for whom advanced disease rates are up to 4-fold higher, and may reflect an earlier onset of disease by approximately 5 years. The fact that a racial disparity exists has long been recognized, but the explanations for it remain poorly understood. Potential roots of the disparity include genetic, environmental, and access to care-related factors—and it is almost certain that all three types of factors are partially responsible. African American men are slightly more likely to be screened at younger ages, and less likely to be screened at older ages, compared to Caucasian men. However, overall rates of screening for younger African American men remain too low. The literature is conflicting as to whether African American men are treated differently after diagnosis; in most analyses, race drops out of most multivariable models once other sociodemographic parameters are included. Likewise, raise drops out of most multivariable prognostic models of outcomes, suggesting that while African American men have more aggressive disease on average, this discrepancy is, for the most part, adequately reflected in risk factors like Gleason score and stage. More intensive use of PSA testing at younger ages for African American men would likely go a long way toward narrowing the mortality gap; future research is needed to understand the interactions between potentially modifiable genomic and environmental factors in order to adequately ameliorate the disparity.

Learning Objectives:

1. Understand the evolving epidemiology of prostate cancer in the the US and globally, and the excess burden of disease borne by men of African ancestry

2. Describe the factors, general and specific, that may underlie and explain the racial disparity.

Learning Objectives:

1. To provide an overview of relevant hereditary cancer syndromes

2. To encourage providers to obtain a thorough family history

3. To educate providers about available risk assessment strategies for the identification of patients at risk for hereditary cancer syndromes as set forth in current guidelines

4. To inform providers about the spectrum of available genetic tests, the content of genetics consultations, and the meaning and implications of genetic test results

5. To guide providers about when to refer to a genetics professional and/or other specialists as needed

Cancer disparities among persons of African descent are driven by both biological and nonbiological factors. There is evidence in breast cancer that psychosocial factors (environment, socioeconomic status, health behaviors, etc.) have a strong influence on racial mortality. After controlling for these factors, overall phenotypic differences in breast cancer pathology remain even after consideration of geographic ancestry. Current molecular evidence suggests that chronic/reoccurring inflammation, driven I in part by the innate immune pathways, contribute to cancer progression and possible outcomes. It is established that one of the most diverse and least well-characterized genomes in the human family are those individuals of African ancestry. Germline variations in innate immune genes have been retained in the human genome offer enhanced protection against environmental pathogens, and protective innate immune variants against specific pathogens.  We will interrogate the role of innate immune variants as part of the basis for explaining the biology of cancer health disparities of persons of African and European ancestry. Evidence will be presented that suggests that racial/ethnic differences in innate immune programs will explain the ethnic differences in both pro- and antitumor immunity, tumor progression, and prognosis, supporting the phenomenon of racial/ethnic disparities in cancer. This presentation will explore examples of protective innate immune genetic variants that are distributed disproportionately among racial populations and linked with racial/ ethnic disparities of breast and prostate cancer.

Learning Objectives:

1. At the conclusion of this presentation, participants will be able to discuss the role of geographical ancestry and breast and prostate cancer disparities in persons of African descent

2. Participants will assess the role of innate immune variants in cancer disparities in persons of African descent

Changes in transcriptional regulation are thought to be one of the key drivers of carcinogenesis. Although next-generation sequencer revolutionized transcriptome analysis, there are limitations in the analysis of full-length transcripts with short-read sequencing data. We developed a multi-sample long-read transcriptome assembly pipline, MuSTA, and showed through simulation that a transcriptome can be constructed from the transcripts represented by the target samples, enabling accurate evaluation of transcriptional regulation. RNA extracted from 22 breast cancer clinical specimens were subjected to Iso-seq full-length transcriptome sequencing using Sequel (PacBio). The MuSTA pipeline was applied to the long-read sequencing data to successfully obtain a full-length transcriptome for the entire cohort. By comparing isoform existence and expression between estrogen receptor positive and triple-negative subtypes, we obtained a comprehensive set of subtype-specific isoforms and differentially used isoforms which consisted of both known and unannotated isoforms. We also found that the exon-intron structure of fusion transcripts depends on the features of the involved genomic regions, and that three-piece fusion transcripts were transcribed from complex structural variations.

Learning Objectives:

1. To understand the somatic structural variations of cancer genomes

2. To understand the technical difficulties in transcript analysis with long-read sequencers

3. To find out how transcriptome analysis using a long-read sequencer is useful for understanding cancer biology

Background: Healthcare access (HCA) encompasses five important dimensions: Affordability, Availability, Accessibility, Accommodation and Acceptability. Published studies of cancer typically focus on affordability, availability and accessibility, however accessibility and accommodation are typically less studied

Objectives: The goal of the current study is to conduct a qualitative study characterizing cancer survivor’s perceptions of HCA dimensions and to identify key facilitators or barriers to equitable healthcare among Black, Hispanic and White cancer survivors

Methods: We recruited a total of 32 cancer survivors into a qualitative study to understand their cancer treatment journey and experiences accessing healthcare. Over 60% of participants were Black, and 18% were Hispanic. Focus groups were moderated by an experienced facilitator, and data were coded and analyzed using NVivo 12 to identify key themes. Peer debriefing was done to evaluate the credibility and completeness of analysis

Results: Across the five HCA dimensions evaluated, acceptability emerged as the most important among cancer survivors. Acceptability was discussed in a variety of positive ways, such as the importance of bedside manner, patient-provider communication, and empathy. Accommodation—relating to ability to see provider when convenient, limited language barriers—were also identified as important. Faith and access to support groups were noted as facilitators of healthcare access, while role conflict was noted as a barrier

Conclusion: Access to healthcare is a top priority for cancer survivors, and impacts quality of initial treatment, supportive care and survival. Even when there were underlying concerns about affordability and accessibility, issues related to acceptability were key to survivor’s feeling a sense of psychological safety with their providers might be a specific, quantifiable and impactful measure of patient-centered, high-quality care

Learning Objectives:

1. Understand the 5 dimensions of healthcare access

2. Identify the salient healthcare access dimension for cancer survivors

Join Branden Hopkinson, Product Marketing Manager at Visiopharm, who will talk about using AI classifiers to find patterns in your images and which artificial intelligence method will work best with your image data. The terms “artificial intelligence” (AI), “machine learning” (ML), and “deep learning” (DL) have infiltrated nearly every industry. This presentation will define the important differences between each term, provide understanding of their roles in image analysis, and help you to decide which questions to ask and when to employ each method in your research.

Learning Objectives:

1. Explain the terms artificial intelligence, machine learning and deep learning as it relates to image analysis

2. Understand the Artificial Intelligence classifiers that Visiopharm software utilizes

3. Learn the basics of when to use these classifiers to identify different patterns in your data

Cancer is a multifactorial disease, and its genesis and progression are extremely complex. The scope of precision oncology is rapidly expanding to address previously undruggable targets. The term “undruggable” was used to describe proteins that could not be targeted pharmacologically. Many targets in cancer research fall into this category, including KRAS, which has eluded researchers for more than 30 years until recent landmark discoveries of AMG 510, MRTX 849, and BI 1701963. As clinical trials are vigorously advanced, efficacy, safety, resistance, and combination strategies are being carefully evaluated for those drug candidates. In addition to KRAS, there are other challenging targets such as P53, MYC, fusion transcription factors, immuno-suppressors,…etc. The focus of this webinar is to provide an overview of strategies in drugging some perceived undruggable targets, with stories and case studies by scientists who strive to find these medicines and fill unmet patient needs. Experienced in overcoming prior hurdles to find effective antagonists against cancer, as a R&D CRO, ChemPartner has utilized a wide range of integrated approaches such as medicinal chemistry, antibody-drug conjugates, monoclonal antibodies, in vitro biology assays, cell biology, immuno-oncology, pharmacology, and combination therapy to identify and tackle the problems that surround drug development for “undruggable” targets.

Tumor phenotypes are dictated not only by the neoplastic cell component, but also by the tumor microenvironment (TME), which is inherently immuno-suppressive, is equipped to hamper effector T-cell function and includes immune and inflammatory cells, soluble mediators such as interferon (IFN)-gamma and extracellular matrix components. Acute myeloid leukemia (AML) is characterized by clonal expansion of poorly differentiated myeloid precursors, resulting in impaired hematopoiesis and often bone marrow (BM) failure. TP53 mutations occur in 8-10% of de novo AML and are associated with chemotherapy refractoriness and with poor prognostic features. Our multi-institutional study was undertaken to characterize the immune ecosystem of non-promyelocytic AML with TP53 mutations using the nCounter™ system (NanoString Technologies Inc., Seattle, WA), with the ultimate goal to implement new immunotherapy agents for patients harboring this molecularly defined subtype of AML. We detected high T-cell infiltration and high expression of immune checkpoints and IFN- signaling molecules in patients with TP53 mutated AML compared with AML subgroups with other risk-defining molecular lesions. We also computed an experimentally derived, TP53-related immune gene signature which stratified survival in a broad cohort of TCGA AML cases. Finally, our correlative analyses in patients with relapsed/refractory AML treated with flotetuzumab, an investigational, bispecific CD123×CD3 DART® molecule, showed therapeutic efficacy in individuals with altered TP53 status and identified immune gene signatures that support the prediction of clinical responses. In conclusion, our study has identified unique immunological profiles in patients with TP53 mutated AML. From a clinical standpoint, ‘immune enriched’ AMLs might be amenable to immunotherapy approaches with T-cell engagers.

Learning Objectives:

1. Learn about unique immunological profiles of patients with acute myeloid leukemia (AML)

2. Explore protein signatures and how they influence immunotherapy treatment choices

3. Learn how digital spatial profiling is impacting the field of oncology

Robotic partial nephrectomy (RPN) has increasingly been applied to the treatment of small renal masses. Because of space limitations and the size of the robot at the patient side, the standard approach to RPN has been transperitoneal.  However, posterior located tumors are difficult to approach transperitoneally, and require the kidney to be completely mobilized and flipped medially.    Retroperitoneal robotic partial nephrectomy is an alternative technique ideally suited for posterior located tumors and for patients who have had previous surgery.  The approach eliminates the need to rotate the kidney and avoids manipulation of the intestines.  Consequently, patients are able to eat and leave the hospital sooner in our experience.  The evolution of the daVinci robot has facilitated the retroperitoneal approach to partial nephrectomy by allowing the robotic arms to be placed closer and permitting more range of movement. Retroperitoneal robotic partial nephrectomy is a safe and reproducible approach to minimally invasive partial nephrectomy.

Learning Objectives:

1. Understand the indications for retroperitoneal robotic partial nephrectomy

2. Identify the retroperitoneal landmarks, port configuration and docking for retroperitoneal robotic partial nephrectomy

3. Understand the techniques for creating and optimizing the retroperitoneal space

This webinar covers our latest solutions for improved culture of primary and xenotransplanted tumor cells. Get insights into dissociation of fresh tumor samples into viable single-cell suspensions, untouched isolation of fully viable, heterogeneous tumor cells, and in vitro derivation and expansion of primary and xenotransplanted tumor cell cultures while preserving parental tumor heterogeneity, tumor-initiating capacity, and genetic stability. Learn about our latest solutions for in vitro derivation and expansion of primary and xenotransplanted tumor cell cultures while preserving parental tumor heterogeneity, tumor-initiating capacity, and genetic stability.

Learning Objectives:

1. Learn how you can improve your in vitro models for cancer research

2. Get tips and tricks for maintaining primary tumor cell cultures

3. See the latest advances in tumor cell isolation

This lecture will encompass the learning experience I have had over my 14,000 RALP cases.  We will present an evidence based prospective data collection and analysis of RALP outcomes.  We will describe evolution of technique for continence, potency and cancer control. Each technical innovation has led to an improvement in overall outcome.  New technologies such as single port robots have also now entered the picture and are presenting a new learning curve.

Learning Objectives:

1. Understand the key outcome metrics for RALP

2. Understand the technical nuances of the operation

3. Analyze the key patient factors that influence RALP outcomes

Immune therapy approaches are being used in many different solid cancers today including clear cell renal carcinoma (mccRCC) and metastatic prostate adenocarcinoma. Over the past few years immune therapy has moved from later stage therapy in mccRCC to first line therapy. Additionally, combination therapy appears to be more effective than single-agent immune therapy in this disease setting. This presentation will compare the combination first-line therapies for mccRCC. Also we will discuss the current role for immune therapy in mPC and the ideal patient to treat with immune therapy. Finally we will discuss possible future combination immune therapies for mPC.

Learning Objectives:

1. Understand the optimal first-line therapies for patients with metastatic clear cell renal carcinoma

2. Understand the impact of cancer vaccines in the treatment of metastatic testosterone suppression resistant prostate cancer (mCRPC)

3. Define the optimal patient likely to benefit from Provenge

Activating mutations in PIK3CA are frequent in human breast cancer, and phosphoinositide 3-kinase alpha (PI3Kα) inhibitors have been approved for therapy. To characterize determinants of sensitivity to these agents, we analyzed PIK3CA-mutant cancer genomes and observed the presence of multiple PIK3CA mutations in 12 to 15% of breast cancers and other tumor types, most of which (95%) are double mutations. Double PIK3CA mutations are in cis on the same allele and result in increased PI3K activity, enhanced downstream signaling, increased cell proliferation, and tumor growth. The biochemical mechanisms of dual mutations include increased disruption of p110α binding to the inhibitory subunit p85α, which relieves its catalytic inhibition, and increased p110α membrane lipid binding. Double PIK3CA mutations predict increased sensitivity to PI3Kα inhibitors compared with single-hotspot mutations.

Learning Objectives:

1. Understand the indications for PI3K inhibitors in breast cancer

2. Discuss double PIK3CA mutations as a biomarker of increased response to PI3K inhibitors in breast cancer