This document summarizes Erik Rogers' comprehensive exam focusing on hypoxia-driven induction of M2 tumor-associated macrophages. It discusses cancer statistics, the epithelial-to-mesenchymal transition process in cancer progression, and interactions between tumor cells, macrophages, and T-helper cells in the tumor microenvironment. The document outlines two specific aims investigating whether hypoxia and HIF-1 regulate the M2 phenotype and whether M2 macrophages contribute to EMT and invasiveness through MMP production. Experiments are proposed to address these aims and alternative approaches are discussed.
1) CAR T cells specific for CD19 have shown remarkable success in treating blood cancers but have had limited effect on solid tumors which lack the CD19 antigen.
2) The IMPACT technology uses fusion proteins containing the extracellular domain of CD19 linked to single-chain antibodies targeting other antigens to bridge CAR19 T cells to solid tumors.
3) Experiments showed CAR19 T cells redirected by a CD19-anti-HER2 fusion protein were cytotoxic against HER2-positive ovarian cancer cells in vitro, demonstrating the IMPACT technology can redirect CAR T cells to new tumor targets through antigen bridging.
Slide deck used for video presentation to the OCTS conference. Her2-positive CNS metastases plague refractory breast cancer patients. Here we present our wholly novel approach to cell therapy for the treatment of these patients. #celltherapy #oncology #breastcancer
Silencing c-Myc translation as a therapeutic strategy through targeting PI3Kd...Mark Lipstein
This document summarizes a study examining the combination of a novel PI3Kδ inhibitor, TGR-1202, with the proteasome inhibitor carfilzomib for treating hematological malignancies. The study found that TGR-1202 synergizes strongly with carfilzomib in lymphoma, leukemia, and myeloma cell lines and primary cells by silencing c-Myc translation. This synergistic effect is driven by TGR-1202's unexpected additional activity of inhibiting CK1ε, which contributes to repressing phosphorylation of 4E-BP1 and lowering c-Myc protein levels. The results suggest that TGR-1202, as a dual PI3Kδ/CK1ε inhibitor, may have
The document provides information about epithelial-mesenchymal transition (EMT) and its role in development, disease, and signaling pathways. It discusses:
1) The differences between epithelial and mesenchymal cell markers and properties. 2) How EMT converts epithelial cells into mesenchymal cells through disrupting cell adhesion and interactions with the extracellular matrix. 3) The transient and context-specific nature of EMT. 4) The role of EMT in embryogenesis, cancer progression and metastasis, and fibrosis. 5) Signaling pathways and factors that activate and suppress EMT such as TGF-β, Wnt, Notch, hypoxia, and inflammation. 6) How EMT generates cancer stem-like cells and
1. The study found that ADAM8, a protein highly expressed in triple-negative breast cancers, regulates the expression of miRNAs, including miR-720.
2. Experiments showed that ADAM8 induces miR-720 expression via activation of the β1-integrin/ERK signaling pathway.
3. Modulating miR-720 levels in triple-negative breast cancer cells revealed that miR-720 promotes migratory and invasive abilities, suggesting it plays a role in the aggressive phenotype driven by ADAM8.
Maytansinoid immunoconjugate IMGN901 is cytotoxicEllen Gunn
IMGN901 is a maytansinoid immunoconjugate targeting CD56, which is expressed in 70% of multiple myeloma cases. The study evaluated IMGN901 alone and in combination with lenalidomide and dexamethasone to determine if it can overcome environmental-mediated drug resistance (EM-DR) in multiple myeloma. While adhesion to extracellular matrix reduced IMGN901 potency, it remained cytotoxic with an LC50 of 43 nM. However, only a combination of IMGN901, lenalidomide, and dexamethasone was able to overcome drug resistance arising from direct contact between multiple myeloma and stromal cells. Drug resistance in this condition was associated with upregulation of multi-drug
This document discusses regulation of deoxynucleotide metabolism in cancer and its therapeutic implications. It describes how imbalanced levels of deoxynucleotide triphosphates (dNTPs) can lead to genomic instability and increased cancer risk. Several key points are made: 1) Enzymes like ribonucleotide reductase and SAMHD1 regulate dNTP levels and maintain genomic stability. 2) Mutations or dysregulation of these enzymes can cause elevated dNTP pools and increased mutagenesis, facilitating cancer development. 3) Many cancer therapies target dNTP synthesis pathways to inhibit tumor growth. 4) SAMHD1 specifically acts as a tumor suppressor by maintaining low dNTP levels
1) CAR T cells specific for CD19 have shown remarkable success in treating blood cancers but have had limited effect on solid tumors which lack the CD19 antigen.
2) The IMPACT technology uses fusion proteins containing the extracellular domain of CD19 linked to single-chain antibodies targeting other antigens to bridge CAR19 T cells to solid tumors.
3) Experiments showed CAR19 T cells redirected by a CD19-anti-HER2 fusion protein were cytotoxic against HER2-positive ovarian cancer cells in vitro, demonstrating the IMPACT technology can redirect CAR T cells to new tumor targets through antigen bridging.
Slide deck used for video presentation to the OCTS conference. Her2-positive CNS metastases plague refractory breast cancer patients. Here we present our wholly novel approach to cell therapy for the treatment of these patients. #celltherapy #oncology #breastcancer
Silencing c-Myc translation as a therapeutic strategy through targeting PI3Kd...Mark Lipstein
This document summarizes a study examining the combination of a novel PI3Kδ inhibitor, TGR-1202, with the proteasome inhibitor carfilzomib for treating hematological malignancies. The study found that TGR-1202 synergizes strongly with carfilzomib in lymphoma, leukemia, and myeloma cell lines and primary cells by silencing c-Myc translation. This synergistic effect is driven by TGR-1202's unexpected additional activity of inhibiting CK1ε, which contributes to repressing phosphorylation of 4E-BP1 and lowering c-Myc protein levels. The results suggest that TGR-1202, as a dual PI3Kδ/CK1ε inhibitor, may have
The document provides information about epithelial-mesenchymal transition (EMT) and its role in development, disease, and signaling pathways. It discusses:
1) The differences between epithelial and mesenchymal cell markers and properties. 2) How EMT converts epithelial cells into mesenchymal cells through disrupting cell adhesion and interactions with the extracellular matrix. 3) The transient and context-specific nature of EMT. 4) The role of EMT in embryogenesis, cancer progression and metastasis, and fibrosis. 5) Signaling pathways and factors that activate and suppress EMT such as TGF-β, Wnt, Notch, hypoxia, and inflammation. 6) How EMT generates cancer stem-like cells and
1. The study found that ADAM8, a protein highly expressed in triple-negative breast cancers, regulates the expression of miRNAs, including miR-720.
2. Experiments showed that ADAM8 induces miR-720 expression via activation of the β1-integrin/ERK signaling pathway.
3. Modulating miR-720 levels in triple-negative breast cancer cells revealed that miR-720 promotes migratory and invasive abilities, suggesting it plays a role in the aggressive phenotype driven by ADAM8.
Maytansinoid immunoconjugate IMGN901 is cytotoxicEllen Gunn
IMGN901 is a maytansinoid immunoconjugate targeting CD56, which is expressed in 70% of multiple myeloma cases. The study evaluated IMGN901 alone and in combination with lenalidomide and dexamethasone to determine if it can overcome environmental-mediated drug resistance (EM-DR) in multiple myeloma. While adhesion to extracellular matrix reduced IMGN901 potency, it remained cytotoxic with an LC50 of 43 nM. However, only a combination of IMGN901, lenalidomide, and dexamethasone was able to overcome drug resistance arising from direct contact between multiple myeloma and stromal cells. Drug resistance in this condition was associated with upregulation of multi-drug
This document discusses regulation of deoxynucleotide metabolism in cancer and its therapeutic implications. It describes how imbalanced levels of deoxynucleotide triphosphates (dNTPs) can lead to genomic instability and increased cancer risk. Several key points are made: 1) Enzymes like ribonucleotide reductase and SAMHD1 regulate dNTP levels and maintain genomic stability. 2) Mutations or dysregulation of these enzymes can cause elevated dNTP pools and increased mutagenesis, facilitating cancer development. 3) Many cancer therapies target dNTP synthesis pathways to inhibit tumor growth. 4) SAMHD1 specifically acts as a tumor suppressor by maintaining low dNTP levels
Canine oncoprotein targets for Melanoma, Breast Cancer, OsteosarcomaSnehal Salunkhe
We can acquire the basic knowledge about canine oncoprotein targets specifically for Melanoma, Osteosarcoma and Breast cancer. I haven't mentioned all the proteins and their targets involved, but just a general overview of these targets with the drugs involved in their treatment/clinical trials.
This presentation gives the basic idea, about the information on the role of tyrosine kinases in cancer. I have also included a phylogenetic tree for finding the relatedness between different organisms.
1) The study found higher levels of MT1-MMP and lower levels of RECK, an MT1-MMP inhibitor, on circulating human CD34+ progenitor cells compared to those in bone marrow. MT1-MMP levels correlated with successful mobilization of CD34+ cells in healthy donors and patients receiving G-CSF treatment.
2) Treatment with G-CSF further increased MT1-MMP and decreased RECK expression on human and mouse hematopoietic cells in a PI3K/Akt-dependent manner. Blocking MT1-MMP impaired chemotaxis and homing of mobilized human CD34+ progenitors.
3) Mobilization of human progen
Regulatory T-cells (Tregs) help maintain self-tolerance and prevent autoimmunity by suppressing immune responses. They express FOXP3 and CD25 and function through various mechanisms like secreting inhibitory cytokines or metabolizing IL-2. Tregs are implicated in tumor immune escape by suppressing anti-tumor immunity. While Tregs are normally beneficial, in cancer high levels associate with poor prognosis by hindering immune response. Emerging immunotherapies aim to deplete or modulate Tregs to enhance anti-tumor immunity.
This slideshow gives all the basic information about Canine Mast Cell Tumor such as introduction to mast cells, mast cell tumor, diagnosis, grading of tumors, the immunohistochemistry of the tumors, treatment etc.
The document discusses cancer immunotherapy and biomarkers. It provides diagrams of immune checkpoint blockade showing how CTLA-4 and PD-1 inhibitors work. It lists FDA-approved immune checkpoint inhibitors across different cancer types. Emerging immunotherapy targets and combinations are discussed, as well as current and emerging biomarkers like PD-L1 expression, MSI/MMR status, and tumor mutational burden that can help identify patients most likely to respond to immunotherapy. Practice aids provide more details on mechanisms, targets, and biomarker testing.
- The study examines the expression of COL11A1/procollagen 11A1 in human mesenchymal cells, colon adenocarcinoma cells, and cancer-associated stromal cells.
- Immunostaining showed procollagen 11A1 expression in human bone marrow mesenchymal cells and colon cancer stromal cells, but not in normal colon cells.
- In colon cancers, high procollagen 11A1 expression in stromal cells was associated with nodal involvement, distant metastases, and advanced disease stage.
This document summarizes recent research on the role of noncoding RNAs, such as microRNAs and long noncoding RNAs, in prostate cancer progression and castration-resistant prostate cancer. It discusses how some noncoding RNAs are regulated by the androgen receptor and promote tumor growth by influencing processes like apoptosis, cell cycle, and cell invasion. The expression levels of certain microRNAs have been associated with prostate cancer diagnosis, aggressiveness, and treatment resistance. Integrative analyses of androgen receptor binding sites and regulated transcripts have provided new insights into the complex molecular mechanisms driven by noncoding RNAs in prostate cancer.
This document compares the efficacy of 5 different FGFR inhibitors - AZD4547, BGJ398, JNJ42756493, PD173074 and TKI258 - at inhibiting FGFR1 signaling and cell proliferation in solid tumor cell lines and leukemia cell lines that overexpress FGFR1. It finds that the pan-FGFR inhibitors AZD4547, BGJ398 and JNJ42756493 more potently suppress FGFR1 phosphorylation and downstream signaling, cause more cell cycle arrest, and have lower GI50 values, indicating they more efficiently inhibit cell growth. It also shows that BGJ398 treatment significantly suppresses leukemia progression in a xenograft model of primary human AML overexpressing FGFR1, supporting
The document discusses immunotherapy and the role of pathologists in assessing tumor samples. It describes how certain tumors express PD-L1 antigens that can be recognized by the immune system, but the tumors also engage immune checkpoint pathways like PD-1 and CTLA-4 to evade the immune response. Immunotherapy drugs target these checkpoint pathways to enhance the immune response. The document outlines the FDA-approved PD-L1 immunohistochemistry assays and biomarkers used to identify cancer patients most likely to respond to immune checkpoint inhibitors for various cancer types including NSCLC, melanoma, bladder cancer, and colorectal cancer.
Fisetin, a naturally occurring flavonoid, potentiates the effects of sorafenib, a RAF inhibitor, in reducing the growth of BRAF-mutated melanoma cells. Combination treatment of fisetin and sorafenib more effectively inhibited cell proliferation, induced apoptosis, and reduced colony formation at lower doses compared to either agent alone. The combination treatment also more strongly reduced tumor growth in mouse models of melanoma compared to monotherapies. The enhanced effects are attributed to simultaneous inhibition of the MAPK and PI3K signaling pathways, which play key roles in melanoma cell survival and proliferation.
This document discusses recent advances in paediatric oncology and cancer immunotherapy. It provides details on adaptive cell transfer therapy including TIL therapy, TCR therapy and CAR-T cell therapy. It summarizes key studies demonstrating the regression of cancer using TILs and the use of T cells transduced with TCRs or CARs. It also outlines the structure and components of CARs including antigen targeting domains, costimulatory domains and signalling domains. The document discusses clinical trials demonstrating high response rates in ALL patients treated with CD19-targeted CAR T-cell therapy. It summarizes treatment approaches and outcomes for different subtypes of ALL including Ph-like ALL.
Basic Mutagenic signal Transduction or the cancer signal transduction that control cell cycle are important pathways to understand cancer in molecular level and to invent targeted treatment.
This document discusses fibroblast growth factors (FGFs) and their receptors (FGFRs) in cancer. It notes that FGFRs are cell surface receptors involved in normal cellular processes but also cancer development and progression. FGFRs may promote cancer cell growth, invasion, and angiogenesis. The document provides details on FGF signaling pathways, mutations in FGFR genes linked to various cancers, and differential expression of FGFs/FGFRs across cancer types like breast, bladder, prostate, lung and multiple myeloma. It concludes that deregulation of FGFs/FGFRs supports cancer progression and discusses challenges in targeting them for cancer therapy.
Identification of personalized therapies for LKB1 Mutant lung cancer using a ...Alex Yang
1) Researchers performed a high throughput screen of FDA-approved mitochondrial inhibitors to identify compounds that selectively kill LKB1 mutant lung cancer cells.
2) They identified MI007 as an inhibitor that induces loss of mitochondrial membrane potential and apoptosis in LKB1 mutant cells at low, clinically relevant doses but not in wild-type cells.
3) Further experiments showed that MI007 induces markers of apoptosis, energy stress, and mitochondrial stress in LKB1 mutant mouse and human lung cancer cell lines and mouse lung tumors. Researchers conclude that MI007 and other mitochondrial inhibitors warrant further preclinical testing for treating LKB1 mutant lung cancer.
The document describes research showing that treatment with SIN3 interaction domain (SID) decoy peptides inhibits invasion and Wnt/β-catenin signaling in triple-negative breast cancer cells. The SID decoy peptides bind to the PAH2 domain of SIN3A, disrupting its interaction with the transcription factor TGIF1. TGIF1 knockdown also inhibited Wnt target genes and cell invasion. The findings suggest targeting the SIN3A-TGIF1 interaction with SID decoys is a novel strategy to block tumor invasion and metastasis by reversing the epithelial-to-mesenchymal transition program and inhibiting Wnt/β-catenin signaling in triple-negative breast cancer.
1) GRK5 regulates prostate cancer cell migration and invasion in vitro and tumor growth and metastasis in vivo.
2) GRK5 phosphorylates the cytoskeletal protein moesin, regulating its subcellular distribution and localization to the cell periphery.
3) Phosphorylation of moesin at threonine 66 by GRK5 is important for cell spreading, and mutation of this site reduces cell spreading.
Hallmarks of cancer and radiopharmaceuticalsAlice Viana
In this presentation I review the article Hallmarks of cancer: next generation, from Hanahan and Weinberg, and make a parallel with potential and current targets of radiopharmaceuticals for diagnosis and treatment.
Macrophage polarization refers to the process by which macrophages diversify and take on different phenotypes in response to signals from the tumor microenvironment. The two main polarization states are M1 and M2 macrophages. M1 macrophages have anti-tumor properties while M2 macrophages, also known as tumor-associated macrophages (TAMs), promote tumor progression. TAMs influence processes like angiogenesis, immune suppression, tumor growth, metastasis, and chemotherapy resistance. Targeting TAMs is a promising strategy for cancer treatment, and several drugs aim to reprogram TAMs from the M2 phenotype to the M1 phenotype or directly kill M2 macrophages.
Canine oncoprotein targets for Melanoma, Breast Cancer, OsteosarcomaSnehal Salunkhe
We can acquire the basic knowledge about canine oncoprotein targets specifically for Melanoma, Osteosarcoma and Breast cancer. I haven't mentioned all the proteins and their targets involved, but just a general overview of these targets with the drugs involved in their treatment/clinical trials.
This presentation gives the basic idea, about the information on the role of tyrosine kinases in cancer. I have also included a phylogenetic tree for finding the relatedness between different organisms.
1) The study found higher levels of MT1-MMP and lower levels of RECK, an MT1-MMP inhibitor, on circulating human CD34+ progenitor cells compared to those in bone marrow. MT1-MMP levels correlated with successful mobilization of CD34+ cells in healthy donors and patients receiving G-CSF treatment.
2) Treatment with G-CSF further increased MT1-MMP and decreased RECK expression on human and mouse hematopoietic cells in a PI3K/Akt-dependent manner. Blocking MT1-MMP impaired chemotaxis and homing of mobilized human CD34+ progenitors.
3) Mobilization of human progen
Regulatory T-cells (Tregs) help maintain self-tolerance and prevent autoimmunity by suppressing immune responses. They express FOXP3 and CD25 and function through various mechanisms like secreting inhibitory cytokines or metabolizing IL-2. Tregs are implicated in tumor immune escape by suppressing anti-tumor immunity. While Tregs are normally beneficial, in cancer high levels associate with poor prognosis by hindering immune response. Emerging immunotherapies aim to deplete or modulate Tregs to enhance anti-tumor immunity.
This slideshow gives all the basic information about Canine Mast Cell Tumor such as introduction to mast cells, mast cell tumor, diagnosis, grading of tumors, the immunohistochemistry of the tumors, treatment etc.
The document discusses cancer immunotherapy and biomarkers. It provides diagrams of immune checkpoint blockade showing how CTLA-4 and PD-1 inhibitors work. It lists FDA-approved immune checkpoint inhibitors across different cancer types. Emerging immunotherapy targets and combinations are discussed, as well as current and emerging biomarkers like PD-L1 expression, MSI/MMR status, and tumor mutational burden that can help identify patients most likely to respond to immunotherapy. Practice aids provide more details on mechanisms, targets, and biomarker testing.
- The study examines the expression of COL11A1/procollagen 11A1 in human mesenchymal cells, colon adenocarcinoma cells, and cancer-associated stromal cells.
- Immunostaining showed procollagen 11A1 expression in human bone marrow mesenchymal cells and colon cancer stromal cells, but not in normal colon cells.
- In colon cancers, high procollagen 11A1 expression in stromal cells was associated with nodal involvement, distant metastases, and advanced disease stage.
This document summarizes recent research on the role of noncoding RNAs, such as microRNAs and long noncoding RNAs, in prostate cancer progression and castration-resistant prostate cancer. It discusses how some noncoding RNAs are regulated by the androgen receptor and promote tumor growth by influencing processes like apoptosis, cell cycle, and cell invasion. The expression levels of certain microRNAs have been associated with prostate cancer diagnosis, aggressiveness, and treatment resistance. Integrative analyses of androgen receptor binding sites and regulated transcripts have provided new insights into the complex molecular mechanisms driven by noncoding RNAs in prostate cancer.
This document compares the efficacy of 5 different FGFR inhibitors - AZD4547, BGJ398, JNJ42756493, PD173074 and TKI258 - at inhibiting FGFR1 signaling and cell proliferation in solid tumor cell lines and leukemia cell lines that overexpress FGFR1. It finds that the pan-FGFR inhibitors AZD4547, BGJ398 and JNJ42756493 more potently suppress FGFR1 phosphorylation and downstream signaling, cause more cell cycle arrest, and have lower GI50 values, indicating they more efficiently inhibit cell growth. It also shows that BGJ398 treatment significantly suppresses leukemia progression in a xenograft model of primary human AML overexpressing FGFR1, supporting
The document discusses immunotherapy and the role of pathologists in assessing tumor samples. It describes how certain tumors express PD-L1 antigens that can be recognized by the immune system, but the tumors also engage immune checkpoint pathways like PD-1 and CTLA-4 to evade the immune response. Immunotherapy drugs target these checkpoint pathways to enhance the immune response. The document outlines the FDA-approved PD-L1 immunohistochemistry assays and biomarkers used to identify cancer patients most likely to respond to immune checkpoint inhibitors for various cancer types including NSCLC, melanoma, bladder cancer, and colorectal cancer.
Fisetin, a naturally occurring flavonoid, potentiates the effects of sorafenib, a RAF inhibitor, in reducing the growth of BRAF-mutated melanoma cells. Combination treatment of fisetin and sorafenib more effectively inhibited cell proliferation, induced apoptosis, and reduced colony formation at lower doses compared to either agent alone. The combination treatment also more strongly reduced tumor growth in mouse models of melanoma compared to monotherapies. The enhanced effects are attributed to simultaneous inhibition of the MAPK and PI3K signaling pathways, which play key roles in melanoma cell survival and proliferation.
This document discusses recent advances in paediatric oncology and cancer immunotherapy. It provides details on adaptive cell transfer therapy including TIL therapy, TCR therapy and CAR-T cell therapy. It summarizes key studies demonstrating the regression of cancer using TILs and the use of T cells transduced with TCRs or CARs. It also outlines the structure and components of CARs including antigen targeting domains, costimulatory domains and signalling domains. The document discusses clinical trials demonstrating high response rates in ALL patients treated with CD19-targeted CAR T-cell therapy. It summarizes treatment approaches and outcomes for different subtypes of ALL including Ph-like ALL.
Basic Mutagenic signal Transduction or the cancer signal transduction that control cell cycle are important pathways to understand cancer in molecular level and to invent targeted treatment.
This document discusses fibroblast growth factors (FGFs) and their receptors (FGFRs) in cancer. It notes that FGFRs are cell surface receptors involved in normal cellular processes but also cancer development and progression. FGFRs may promote cancer cell growth, invasion, and angiogenesis. The document provides details on FGF signaling pathways, mutations in FGFR genes linked to various cancers, and differential expression of FGFs/FGFRs across cancer types like breast, bladder, prostate, lung and multiple myeloma. It concludes that deregulation of FGFs/FGFRs supports cancer progression and discusses challenges in targeting them for cancer therapy.
Identification of personalized therapies for LKB1 Mutant lung cancer using a ...Alex Yang
1) Researchers performed a high throughput screen of FDA-approved mitochondrial inhibitors to identify compounds that selectively kill LKB1 mutant lung cancer cells.
2) They identified MI007 as an inhibitor that induces loss of mitochondrial membrane potential and apoptosis in LKB1 mutant cells at low, clinically relevant doses but not in wild-type cells.
3) Further experiments showed that MI007 induces markers of apoptosis, energy stress, and mitochondrial stress in LKB1 mutant mouse and human lung cancer cell lines and mouse lung tumors. Researchers conclude that MI007 and other mitochondrial inhibitors warrant further preclinical testing for treating LKB1 mutant lung cancer.
The document describes research showing that treatment with SIN3 interaction domain (SID) decoy peptides inhibits invasion and Wnt/β-catenin signaling in triple-negative breast cancer cells. The SID decoy peptides bind to the PAH2 domain of SIN3A, disrupting its interaction with the transcription factor TGIF1. TGIF1 knockdown also inhibited Wnt target genes and cell invasion. The findings suggest targeting the SIN3A-TGIF1 interaction with SID decoys is a novel strategy to block tumor invasion and metastasis by reversing the epithelial-to-mesenchymal transition program and inhibiting Wnt/β-catenin signaling in triple-negative breast cancer.
1) GRK5 regulates prostate cancer cell migration and invasion in vitro and tumor growth and metastasis in vivo.
2) GRK5 phosphorylates the cytoskeletal protein moesin, regulating its subcellular distribution and localization to the cell periphery.
3) Phosphorylation of moesin at threonine 66 by GRK5 is important for cell spreading, and mutation of this site reduces cell spreading.
Hallmarks of cancer and radiopharmaceuticalsAlice Viana
In this presentation I review the article Hallmarks of cancer: next generation, from Hanahan and Weinberg, and make a parallel with potential and current targets of radiopharmaceuticals for diagnosis and treatment.
Macrophage polarization refers to the process by which macrophages diversify and take on different phenotypes in response to signals from the tumor microenvironment. The two main polarization states are M1 and M2 macrophages. M1 macrophages have anti-tumor properties while M2 macrophages, also known as tumor-associated macrophages (TAMs), promote tumor progression. TAMs influence processes like angiogenesis, immune suppression, tumor growth, metastasis, and chemotherapy resistance. Targeting TAMs is a promising strategy for cancer treatment, and several drugs aim to reprogram TAMs from the M2 phenotype to the M1 phenotype or directly kill M2 macrophages.
The document summarizes the role of innate and adaptive immune cells in the tumor microenvironment and their effect on tumor growth. It discusses how the tumor microenvironment can influence immune cells and how immune cells can affect tumor progression. Key cells discussed include macrophages, neutrophils, NK cells, T cells, B cells, dendritic cells, and regulatory T cells. It covers topics like hypoxia, inflammation, immune evasion mechanisms used by tumors, and the pro-tumoral phenotypes that immune cells can adopt in the microenvironment.
Cancer Epigenetics: Concepts, Challenges and PromisesMrinmoy Pal
The presentation highlights how recent investigations have shown extensive reprogramming of almost every component of the epigenetic machinery in cancer leading to the emergence of the promising field of epigenetic therapy.
This document discusses molecular approaches and updates in the classification of thyroid neoplasms. It covers:
1. Changes in the WHO classification, emphasizing a molecular-based classification of thyroid tumors including follicular cell-derived and C-cell derived neoplasms.
2. Key driver mutations in different thyroid tumors including BRAF, RAS, RET, NTRK, and others.
3. The importance of immunohistochemistry and molecular testing to classify tumors and guide treatment. Tests for BRAF, RAS, TRK, ALK, and PTEN are highlighted.
4. Specific tumor types are discussed like papillary thyroid carcinoma, follicular thyroid adenoma and carcinoma, and
Cancer stem cells (CSCs) are a small subpopulation of cells within tumors that drive tumor growth and metastasis. They express cell surface markers like CD44 and CD133 and have the ability to self-renew and differentiate. CSCs are resistant to chemotherapy and radiation therapy due to their slow proliferation rate, expression of drug efflux pumps, and increased DNA damage response. Epithelial-mesenchymal transition (EMT) and signaling pathways like TGF-β, WNT and NOTCH regulate CSC properties. Targeting CSCs, their microenvironment, and these pathways may help overcome therapy resistance and prevent cancer recurrence. Identification of CSCs involves flow cytometry analysis of surface markers, dye efflux assays,
1. LAM cells alter monocyte activation and maturation by secreting growth factors that skew macrophages toward a more immature phenotype. Mouse LAM cells decrease nitric oxide production in mouse monocytes and increase expression of genes associated with alternative activation.
2. Human LAM cells also induce an immature phenotype in human monocytes and increase expression of macrophage activation markers. They stimulate chemotaxis of human monocytes through growth factor secretion like VEGF-D.
3. LAM cells likely contribute to inflammatory lung damage in LAM patients by altering monocyte recruitment and maturation through paracrine signaling of growth factors.
Immunotoxins are being developed to specifically target and destroy cancer cells. The document summarizes research from Professor Rama Verma's lab on developing humanized immunotoxins as alternatives to bacterial/toxin-based immunotoxins that can cause toxicity. The lab has created immunotoxins linking GM-CSF to DFF40 to target leukemia cells and IL2 to TRAIL to target leukemia patients' samples. These humanized immunotoxins induce apoptosis in cancer cells with less toxicity than traditional immunotoxins. The lab is also exploring targeting cancer stem cells, which can cause cancer recurrence after treatment.
1. Cancer epigenetics involves heritable changes in gene expression that are not due to changes in DNA sequence. Histone modifications and chromatin remodeling complexes play important roles in cancer development by regulating gene expression and transcription.
2. Many genes that encode histone modifying enzymes are mutated in cancer. Mutations in DNA methyltransferases, histone methyltransferases, and histone demethylases commonly occur in cancers.
3. Targeting epigenetic enzymes and pathways, such as with DNA methyltransferase or histone deacetylase inhibitors, shows promise as cancer therapies. Combination epigenetic and conventional chemotherapy may help reduce drug resistance.
Advanced tumor immunology prof dr.ihsan edan alsaimary university of basrah...dr.Ihsan alsaimary
This document discusses various topics related to tumors and cancer, including:
1. The properties of cancer cells such as being clonally derived, having altered growth patterns, tissue affinities, and chromosomal abnormalities.
2. The development of tumor antigens, including tumor specific antigens from mutations and viral proteins, as well as tumor associated antigens like oncofetal antigens.
3. The process of metastasis, where tumor cells spread via direct seeding, lymphatics, or hematogenous routes, requiring changes in adhesion molecules, proteolytic enzymes and growth factors.
4. Common tumor markers used to screen and monitor cancers, including CEA, CA-125, PSA, and AFP
NIH/NCI/CCR/LCBG Branch Journal Club
Presented Article:
Pencheva N, Tran H, Buss C, Huh D, Drobnjak M, Busam K, Tavazoie SF. Convergent multi-miRNA targeting of ApoE drives LRP1/LRP8-dependent melanoma metastasis and angiogenesis. Cell. 2012 Nov 21;151(5):1068-82.
This document discusses microRNAs (miRNAs) and methods for studying their function and regulation of genes. It describes:
1) What miRNAs are, how they work by incorporating into the RISC complex and repressing target mRNAs through translational repression or degradation.
2) Techniques for manipulating miRNAs in cell lines using reporter assays, mimics, inhibitors and target protectors to study their effects on genes.
3) How to screen for miRNAs that regulate a target gene using ready-made cDNA panels and quantitative PCR. Several examples are provided of identifying miRNAs that regulate important cancer genes.
Metastatic cascade and Epithelial Mesenchymal TransitionShruti Dogra
This document provides an overview of cancer metastasis and the epithelial-mesenchymal transition (EMT) process. It discusses the metastatic cascade, which involves tumor cell invasion, intravasation into blood vessels, transport through circulation, extravasation and homing to distant sites, and formation of secondary tumors. EMT is described as a key step in metastasis that allows epithelial cells to detach from primary tumors and migrate. The molecular and cellular changes involved in EMT include loss of epithelial markers like E-cadherin and gain of mesenchymal markers. Transcription factors such as Snail, Slug, Twist, and ZEB play important roles in inducing EMT. Understanding metastasis and EMT can help develop strategies to prevent cancer spread
This document provides an overview of the host immune response to microbial pathogens, focusing on periodontal bacteria. It discusses how microbe-associated molecular patterns (MAMPs) from bacteria are recognized by pattern recognition receptors (PRRs) like Toll-like receptors (TLRs) and Nucleotide-binding oligomerization domain-like receptors (NLRs). TLR2, TLR4, and TLR9 recognize bacterial lipoproteins, lipopolysaccharide, and CpG-DNA, respectively. NLRs like NOD1 and NOD2 detect peptidoglycan. This interaction stimulates proinflammatory cytokine production and activates innate and adaptive immunity. Recognition of MAMPs is important for the host
This document summarizes molecular studies in central nervous system (CNS) tumors. It discusses the need for molecular studies to integrate histologic and molecular data in tumor classification. It then reviews molecular markers for various CNS tumors including gliomas, medulloblastoma, ependymal tumors, and meningioma. For each tumor type, key genetic alterations are identified that have diagnostic, predictive or prognostic significance such as IDH1/2 mutations, 1p/19q codeletion, MGMT methylation, and others.
This document summarizes a presentation on new targets and agents for hepatocellular carcinoma. It discusses several new targets including VEGFR, PDGFR, c-MET, FGFR4, TGF-β, and PD-1/PDL1 that are being investigated with new agents. Regorafenib and nivolumab are highlighted as agents that have shown survival benefits in late stage trials for HCC. Other agents discussed include tepotinib as a c-MET inhibitor and galunisertib as a TGF-β inhibitor. Combination approaches are also a focus, including using PD-L1 inhibitors with other agents and combining galunisertib with sorafenib.
1) The study investigated differences in microRNA expression profiles between MYC translocation-positive and MYC translocation-negative Burkitt lymphoma cases to uncover potential molecular differences.
2) Unsupervised and supervised analyses revealed distinct microRNA expression patterns between the two groups, with four microRNAs found to be differentially expressed.
3) The different microRNA expression patterns were found to impact gene expression profiles, with DNMT1 and MYCN mRNA expression found to be upregulated in MYC translocation-negative cases. This suggests alternative mechanisms of MYC dysregulation and lymphomagenesis in the absence of translocation.
Characteristic mTOR activity in Hodgkin-lymphomas offers a potential therapeu...Enrique Moreno Gonzalez
Targeting signaling pathways is an attractive approach in many malignancies. The PI3K/Akt/mTOR pathway is activated in a number of human neoplasms, accompanied by
lower overall and/or disease free survival. mTOR kinase inhibitors have been introduced in the therapy of renal cell carcinoma and mantle cell lymphoma, and several trials are currently
underway. However, the pathological characterization of mTOR activity in lymphomas is still incomplete.
This document summarizes theories of carcinogenesis and hallmarks of cancer. It discusses the genetic theory, which states that cancer arises from DNA mutations that are transmitted to daughter cells. It also covers the epigenetic theory, immune surveillance theory, and monoclonal hypothesis. Major hallmarks of cancer include excessive growth from oncogenes, resistance to growth inhibition from tumor suppressor genes like RB and p53, evading apoptosis, angiogenesis, invasion and metastasis. Carcinogenesis is described as a multi-step process involving sequential acquisition of mutations. The roles of growth factors, receptors, signaling proteins, and cell cycle regulators in promoting uncontrolled growth are outlined.
1. Erik Rogers – Comprehensive Exam
08.16.2012
• Hypoxia Driven Induction of M2 Tumor
Associated Macrophages and their
Contributions to EMT and Invasiveness in
Carcinoma Cells
2. 2011 Cancer Statistics
• US: 1.6 million reported cancer cases
• Global: WHO estimates 13.1 million cancer
deaths
• NCI: cancer – “diseases in which abnormal
cells divide without control and are able to
invade other tissues”
4. Epithelial to Mesenchymal Transition
Epithelial cells:
•Apical/Basal polarity
•Selectively permeable barriers
•Anchored to BM
•Cell to cell junctions
Mesenchymal cells:
•Amoeboid movement
•Lamellipodia & Filopodia
•Chemotactic
5. Innate immune response to cancer
• Interactions of 3 cell types are focus of proposal:
– Tumor cells
– Tumor associated macrophages (TAMs: M1 & M2
phenotypes)
– CD4+ T helper cells (TH1 & TH2 phenotypes)
• Carcinoma cells produce cytokines and chemokines that
attract myeloid cells and activate resident myeloid cells
• Naïve T cells (TH0) are induced to TH1 and/or TH2
phenotype by interactions with microenvironment
• Naïve macrophages (Mφ) are induced to M1 and/or M2
phenotype by interactions with microenvironment
6. M1 vs. M2 TAMs
• TH1 cytokines – TNF-α, IFN-γ, IL-1β
– M1 TAM phenotype – inflammatory and cytotoxic
– Primarily localized to invasive front
• TH2 cytokines – IL-4, IL-10, IL-13
– M2 TAM phenotype- immunosuppresive and promote
tumor invasiveness (EMT & metastasis)
– Primarily located intratumorally
• Other microenvironmental activators M2 TAM phenotype
• Hypoxia induces many adaptations in expression patterns
7. Hypoxia M2 TAM TGF-β EMT
• Myeloid Derived Suppressor Cell experiments
– Hypoxia results in MDSCs M2 TAMs
• M2 TAMs co-localize with intraepithelial
fibroblastoid cells
– Correlation between M2 TAM and EMT
• M2 TAMs produce TGF-β
– Snail/Slug, Twist, Smad3/7, PI3K, Notch, GS3Kβ,
NF-ΚB, Hey1, Hes1, etc.
8. HIF-1 Regulation
• Constitutively expressed β-subunit
• α-subunit stabilized in hypoxic conditions
– Normoxia = PHDs 1-3 + pVHL degradation
– PHDs O2 and Fe+2 dependent
– ROS & NO inhibit PHDs oxidize Fe+2 to Fe+3
• 60+ genes regulated by HREs (NCGTG)
– Metabolic adaptation, apoptosis resistance,
angiogenesis and metastasis
9. Model for Hypoxia Driven Feedback
Loop = CD4+TH1
= M1 TAM
= CD4+TH2
= M2 TAM
CCL-2
CXCL-12
CCL-5
CXCL-8
CXCL-12
CCL-2
IL-1
HIF-1
VEGF
EGF
TGFβ
MMP-2
MMP-9IL-4
IL-10
IL-13
10. Specific Aim 1: Questions
• Is hypoxia activated HIF-1 the fundamental
regulator of the induction of Mφ to the M2
phenotype?
• Are M2s required contributors to EMT in
tumor cells?
11. Experimental Design
• Culture M2s in hypoxic conditions –
– Assess for stabilized HIF-1α via Western blot
• Culture wt Mφ in hypoxic conditions –
– Assess for M2 markers via qRT-PCR
• Culture Hif-1αfl/flCre+ Mφ in hypoxic conditions –
– Assess for M2 markers via qRT-PCR
• Co-culture M2s/tumor cells –
– Assess for EMT markers in tumor cells via qRT-PCR
and Immunofluorescence staining
12. Does hypoxia stabilize HIF-1α in M2s?
Mφ M2
--------------β-Actin--------------
•Is hypoxia activated HIF-1 the fundamental
regulator of the induction of the M2
phenotype?
•FACS isolated M2s were cultured
in hypoxic incubator chambers for 4 days
•Mφ were cultured in normoxic conditions
for control comparison
•Western blot analysis indicates presence
of stabilized HIF-1α monomer in M2s
13. Does hypoxia activated HIF-1 induce M2?
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
F4/80 IL-1 IL-6 IL-23 IL-4 IL-10 IL-13
FoldDifference
4 Days in Hypoxia •Is hypoxia activated HIF-1 the
fundamental regulator of the
induction of the M2 phenotype?
•wt Mφ were cultured in hypoxic
incubator chambers for 4 days
•F4/80 pan macrophage marker
set as comparative baseline
•qRT-PCR shows significantly
higher transcription of M2
associated cytokines
14. Does hypoxia induce M2 in
Hif-1αfl/flCre+ Mφs?
0
0.2
0.4
0.6
0.8
1
1.2
F4/80 IL-1 IL-6 IL-23 IL-4 IL-10 IL-13
FoldDifference
4 Days in Hypoxia •Is hypoxia activated HIF-1 the
fundamental regulator of the
induction of the M2 phenotype?
•Hif-1αfl/flCre+ Mφ were cultured in
hypoxic incubator chambers for
4 days
•F4/80 pan macrophage marker
set as comparative baseline
•qRT-PCR shows significantly
higher transcription of M2
associated cytokines
15. M2 TAMs induce ‘Cadherin switch’ in
carcinoma cells
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
E-cadherin N-cadherin vimentin
FoldDifference
EMT Marker Expression
Time 0 48 Hours
•Are M2 TAMs required contributors
to EMT in tumor cells?
•FACS isolated M2 TAMs were co-
cultured with NMuMG cells for 48
hours
•qRT-PCR analysis of mRNA from
sorted NMuMG cells shows down-
regulation of E-cadherin and up-
regulation of N-cadherin and
Vimentin
16. Co-culture of carcinoma cells with M2
TAMs suggests EMT
pMEC cells ---------------E-cadherin-------------- NMuMG cells
pMEC cells --------------N-cadherin--------------- NMuMG cells
•Are M2 TAMs required
contributors to EMT in tumor
cells?
•Co-culture of M2 TAMs with
two different carcinoma cell
lines for 48 hours
•Immunofluorescent staining
Shows down-regulation of
E-cadherin and up-regulation
of N-cadherin
17. Alternative Approaches
• As a means of qualifying the influence of M2
TAMs on NMuMG and pMEC cells, we could
evaluate separate hypoxic cultures of both cell
lines for EMT markers via qRT-PCR.
• Evaluate M2 TAM production of ROS and NO
– 27-dichlorofluorescin diacetate (DCFCA)
• non-fluorescent when reduced
• fluorescent when oxidized
18. Model for Hypoxia Driven Feedback
Loop = CD4+TH1
= M1 TAM
= CD4+TH2
= M2 TAM
CCL-2
CXCL-12
CCL-5
CXCL-8
CXCL-12
CCL-2
IL-1
HIF-1
VEGF
EGF
TGFβ
MMP-2
MMP-9IL-4
IL-10
IL-13
19. Matrix Metalloproteinases
• Three distinct functional domains
– Hemopexin-like C terminal domain
– Catalytic domain
– Pro-domain
• Inactive zymogens via Cys-Zn2+ interaction
– Activated by proteases and/or ROS oxidation
• Activate pro-forms of growth factors
• Release ECM bound growth factors
• Tissue remodeling – EMT, angiogenesis,
metastasis
21. Specific Aim 2: Questions
• Are M1 TAMs polarized to an M2 TAM phenotype in
the tumor microenvironment by IL-4 produced by
CD4+ TH2 lymphocytes?
• Are M2 TAMs indispensable sources of MMP-2 and
MMP-9?
• Are MMP-2 and MMP-9 produced by M2 TAMs
required for tumor invasiveness?
22. Experimental Design
• Evaluate expression of M1 vs. M2 markers after co-culture
of Mφ with CD4+ TH2 cells with and without functional IL-4
• Assess M2 TAM contribution to MMP-2/MMP-9 in tumor
microenvironment
• In vitro invasion assay to assess invasiveness of carcinoma
cells in presence of M2 TAM conditioned media with and
without MMP-2 and MMP-9
• In vivo angiogenesis assay to assess neovascularization in
MMTV-PyMT tumors with and without MMP-2 and MMP-9
23. Is IL-4 critical to induce M1 TAMs to
M2 TAM phenotype?
0 2 4 6 8 10
F4/80
IL-1
IL-6
IL-23
IL-4
IL-10
IL-13
Fold Difference
M1 vs. M2 functional IL-4 activity Are M1 TAMs polarized to M2 TAMs in the
tumor microenvironment by IL-4 produced
by CD4+ TH2 lymphocytes?
4 day co-cultures of M1 TAMs with CD4+ TH2
cells (with and without functional IL-4)
Expression levels of TH1 vs. TH2 cytokines
evaluated in sorted TAMs via qRT-PCR
Sorted TAMs from 4-day co-cultures with
functional IL-4 show predominately TH2
expression pattern
Sorted TAMs from 4-day co-cultures without
functional IL-4 show predominately TH1
expression pattern0 1 2 3 4 5 6
F4/80
IL-1
IL-6
IL-23
IL-4
IL-10
IL-13
Fold Difference
M1 vs. M2 with shRNA Inhibition
of IL-4
24. Are the majority of MMP-2 & MMP-9
produced by M2 TAMs
0.85 0.9 0.95 1 1.05
HCC MMP-2
M2 TAM MMP-2
HCC MMP-9
M2 TAM MMP-9
Fold Difference
Relative MMP Expression
Are M2 TAMs indispensable sources of
MMP-2 and MMP-9?
Expression levels of MMP-2 & MMP-9 were
assessed using RNA isolated from M2 TAM
cultures
Expression levels of MMP-2 & MMP-9 were
assessed using RNA isolated from M2 TAMs
co-cultured with cells obtained from
digested mammary epithelial tumors
Heterogeneous cell cultures (HCC)
The ratio of MMP-2 production in M2 TAMs
compared to HCCs ~1:0.95 and the ratio of
MMP-9 production in M2 TAMs compared
to HCCs ~1:0.93
25. M2 TAM conditioned media – MMPs,
chemoattractants and growth factors
TGF-β EGF VEGF
IL-4 IL-8 IL-10 IL-13 MSF
--------------------------------------------β-actin------------------------------------------
------------------β-actin-----------------
MMP-2 MMP-9
-------------Β-actin-------------
M2 TAM conditioned media in
bottom wells of Boyden Chambers
for in vitro invasion assays
Western blot analysis confirms that
M2 TCM contains cytokines, MMPs
and growth factors
Predicts sufficient stimuli to trigger
invasion of carcinoma cells in upper
wells of Boyden Chamber assays
27. M2 produced MMP-2 / MMP-9
increase angiogenesis
M2/CD4+ TH2 functional MMP2/MMP-9
M2/CD4+ TH2 inhibited MMP2/MMP-9
Are MMP-2 and MMP-9 produced
by M2 TAMs required for tumor
invasiveness?
MMTV-PyMT tumors injected with
M2 TAMs and CD4+ TH2 cells (top).
MMTV-PyMT tumors injected with
M2 TAMs and CD4+ TH2 cells with
inclusion of AG3340 to inhibit
MMP-2/MMP-9 activity (bottom).
Neovascularization assessed by
IF staining for CD-31 (left) and
IHC staining for VEGF:VEGFR on
endothelial cells (right).
28. Alternative Approaches
• Replace M2 TAMs with Mφ in the MMTV-PyMT
tumor injections
• Replace CD4+ TH2 cells with CD8+ cytotoxic T cells in
the MMTV-PyMT tumor injections
• M2 TAMs +/- CD4+ TH2 with ascites fluid from
MMTV-PyMT tumors injected intraperitoneally
and/or sub-cutaneously to assess invasiveness
29. Thank You
• Comprehensive Exam Chair: Dr. Douglas Lake
• Committee Chair: Dr. Alan Rawls
• Committee member: Dr. Jeanne Wilson-Rawls
• Committee member: Dr. Kenro Kusumi
• Thanks to everyone for the time, patience and
guidance