Straightening out the mechanisms of axial elongation using mouse mutant analysis

Understanding the mechanisms that lead to axial elongation in the mouse has direct relevance to elucidating the etiology of vertebral defects in humans. Through the characterization of a spontaneous mouse mutant, kinked tail, and the analysis of Tbx6 protein modulation in vivo, I uncovered two distinct mechanisms affecting axial elongation in the mouse. The kinked tail mutation is a spontaneous mutation, inherited dominantly, that results in a kinky tail phenotype in heterozygotes and early embryonic lethality in homozygotes. Defective axial elongation in kinked tail heterozygotes is displayed as shortened tails and multiple tail kinks resulting from wedge, hemi- and fused vertebrae, similar to those observed in scoliosis patients. These vertebral defects are likely due to a primary notochord defect that is thickened and branched. Kinked tail homozygotes fail to undergo gastrulation due to defective distal visceral endoderm cell migration, ultimately resulting in lethality by embryonic day 8.5. The defective cell migration is further compounded by basement membrane defects and gross dysmorphology of the mutant embryo. Tbx6, a T-box transcription factor, is essential for posterior somite formation, patterning and viability of the mouse embryo. I sought to understand Tbx6 protein regulation and the phenotypic consequences of modulating Tbx6 protein levels in vivo. In vitro analyses revealed that Tbx6 is a relatively stable protein that appears to be regulated in part by the proteasome in addition to other mechanisms. In vivo, less than heterozygous levels of Tbx6 protein results in rib and vertebral defects, enlarged tailbuds and axial shortening while greater than wildtype levels of Tbx6 protein results in small embryonic tailbuds, axial shortening, and lethality. I further examined the consequences of Tbx6 misexpression using a 3-component transgenic system. The primitive streak and presomitic mesoderm are affected in those embryos that misexpress Tbx6.Altogether, my analysis of the spontaneous mutation, kinked tail, demonstrated a possible role of the notochord in proper axial elongation, and the analysis of Tbx6 protein modulation further clarified the importance of maintaining proper levels of Tbx6 for normal axis elongation and embryonic development

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Search for new particles in an extended Higgs sector with four b quarks in the final state at s\sqrt{s} = 13 TeV

A search for an extended Higgs sector, characterized by a massive resonance X decaying to a pair of spin-0 bosons $\phi$ that themselves decay to pairs of bottom quarks, is presented. The analysis is restricted to the mass ranges $m_\phi$ from 25 to 100 GeV and $m_\mathrm{X}$ from 1 to 3 TeV. For these mass ranges, the decay products of each $\phi$ boson are expected to merge into a single large-radius jet. Jet substructure and flavor identification techniques are used to identify these jets. The search is based on CERN LHC proton-proton collision data at $\sqrt{s}$ = 13 TeV, collected with the CMS detector in 2016-2018, corresponding to an integrated luminosity of 138 fb$^{-1}$. Model-specific limits are set on the product of the production cross section and branching fraction for X $\to \phi\phi \to (\mathrm{b\bar{b}})(\mathrm{b\bar{b}})$ as a function of mass, where both the X $\to \phi\phi$ and $\phi \to \mathrm{b\bar{b}}$ branching fractions are assumed to be 100%. These limits are the first of their kind on this process, ranging between 30 and 1 fb at 95% confidence level for the considered mass ranges

Search for new particles in an extended Higgs sector with four b quarks in the final state at s\sqrt{s} = 13 TeV

A search for an extended Higgs sector, characterized by a massive resonance X decaying to a pair of spin-0 bosons $\phi$ that themselves decay to pairs of bottom quarks, is presented. The analysis is restricted to the mass ranges $m_\phi$ from 25 to 100 GeV and $m_\mathrm{X}$ from 1 to 3 TeV. For these mass ranges, the decay products of each $\phi$ boson are expected to merge into a single large-radius jet. Jet substructure and flavor identification techniques are used to identify these jets. The search is based on CERN LHC proton-proton collision data at $\sqrt{s}$ = 13 TeV, collected with the CMS detector in 2016-2018, corresponding to an integrated luminosity of 138 fb$^{-1}$. Model-specific limits are set on the product of the production cross section and branching fraction for X $\to \phi\phi \to (\mathrm{b\bar{b}})(\mathrm{b\bar{b}})$ as a function of mass, where both the X $\to \phi\phi$ and $\phi \to \mathrm{b\bar{b}}$ branching fractions are assumed to be 100%. These limits are the first of their kind on this process, ranging between 30 and 1 fb at 95% confidence level for the considered mass ranges

Search for new particles in an extended Higgs sector with four b quarks in the final state at s\sqrt{s} = 13 TeV

A search for an extended Higgs sector, characterized by a massive resonance X decaying to a pair of spin-0 bosons $\phi$ that themselves decay to pairs of bottom quarks, is presented. The analysis is restricted to the mass ranges $m_\phi$ from 25 to 100 GeV and $m_\mathrm{X}$ from 1 to 3 TeV. For these mass ranges, the decay products of each $\phi$ boson are expected to merge into a single large-radius jet. Jet substructure and flavor identification techniques are used to identify these jets. The search is based on CERN LHC proton-proton collision data at $\sqrt{s}$ = 13 TeV, collected with the CMS detector in 2016-2018, corresponding to an integrated luminosity of 138 fb$^{-1}$. Model-specific limits are set on the product of the production cross section and branching fraction for X $\to \phi\phi \to (\mathrm{b\bar{b}})(\mathrm{b\bar{b}})$ as a function of mass, where both the X $\to \phi\phi$ and $\phi \to \mathrm{b\bar{b}}$ branching fractions are assumed to be 100%. These limits are the first of their kind on this process, ranging between 30 and 1 fb at 95% confidence level for the considered mass ranges

Search for new particles in an extended Higgs sector with four b quarks in the final state at s\sqrt{s} = 13 TeV

A search for an extended Higgs sector, characterized by a massive resonance X decaying to a pair of spin-0 bosons $\phi$ that themselves decay to pairs of bottom quarks, is presented. The analysis is restricted to the mass ranges $m_\phi$ from 25 to 100 GeV and $m_\mathrm{X}$ from 1 to 3 TeV. For these mass ranges, the decay products of each $\phi$ boson are expected to merge into a single large-radius jet. Jet substructure and flavor identification techniques are used to identify these jets. The search is based on CERN LHC proton-proton collision data at $\sqrt{s}$ = 13 TeV, collected with the CMS detector in 2016-2018, corresponding to an integrated luminosity of 138 fb$^{-1}$. Model-specific limits are set on the product of the production cross section and branching fraction for X $\to \phi\phi \to (\mathrm{b\bar{b}})(\mathrm{b\bar{b}})$ as a function of mass, where both the X $\to \phi\phi$ and $\phi \to \mathrm{b\bar{b}}$ branching fractions are assumed to be 100%. These limits are the first of their kind on this process, ranging between 30 and 1 fb at 95% confidence level for the considered mass ranges

Search for new particles in an extended Higgs sector with four b quarks in the final state at s\sqrt{s} = 13 TeV

A search for an extended Higgs sector, characterized by a massive resonance X decaying to a pair of spin-0 bosons $\phi$ that themselves decay to pairs of bottom quarks, is presented. The analysis is restricted to the mass ranges $m_\phi$ from 25 to 100 GeV and $m_\mathrm{X}$ from 1 to 3 TeV. For these mass ranges, the decay products of each $\phi$ boson are expected to merge into a single large-radius jet. Jet substructure and flavor identification techniques are used to identify these jets. The search is based on CERN LHC proton-proton collision data at $\sqrt{s}$ = 13 TeV, collected with the CMS detector in 2016-2018, corresponding to an integrated luminosity of 138 fb$^{-1}$. Model-specific limits are set on the product of the production cross section and branching fraction for X $\to \phi\phi \to (\mathrm{b\bar{b}})(\mathrm{b\bar{b}})$ as a function of mass, where both the X $\to \phi\phi$ and $\phi \to \mathrm{b\bar{b}}$ branching fractions are assumed to be 100%. These limits are the first of their kind on this process, ranging between 30 and 1 fb at 95% confidence level for the considered mass ranges

Search for new particles in an extended Higgs sector with four b quarks in the final state at s\sqrt{s} = 13 TeV

A search for an extended Higgs sector, characterized by a massive resonance X decaying to a pair of spin-0 bosons $\phi$ that themselves decay to pairs of bottom quarks, is presented. The analysis is restricted to the mass ranges $m_\phi$ from 25 to 100 GeV and $m_\mathrm{X}$ from 1 to 3 TeV. For these mass ranges, the decay products of each $\phi$ boson are expected to merge into a single large-radius jet. Jet substructure and flavor identification techniques are used to identify these jets. The search is based on CERN LHC proton-proton collision data at $\sqrt{s}$ = 13 TeV, collected with the CMS detector in 2016-2018, corresponding to an integrated luminosity of 138 fb$^{-1}$. Model-specific limits are set on the product of the production cross section and branching fraction for X $\to \phi\phi \to (\mathrm{b\bar{b}})(\mathrm{b\bar{b}})$ as a function of mass, where both the X $\to \phi\phi$ and $\phi \to \mathrm{b\bar{b}}$ branching fractions are assumed to be 100%. These limits are the first of their kind on this process, ranging between 30 and 1 fb at 95% confidence level for the considered mass ranges