Levels of alternate VEGF ligands in cohort 1 and cohort 2. Error bars represent standard error of the mean.

<p>Following treatment with FOLFIRI and bevacizumab in the discovery cohort, VEGF-C was increased prior to progression and at the time of progression. These findings were similar to previously reported changes from this cohort in PlGF. Levels of VEGF-D were increased at the time of progression, but in contrast to VEGF-C, levels of VEGF-D were not increased prior to progression. In the validation cohort, PlGF was elevated in patients previously receiving chemotherapy and bevacizumab. VEGF-D was also minimally elevated in patients previously receiving chemotherapy and bevacizumab, but elevations in VEGF-C seen in the discovery cohort were not replicated in the validation cohort.</p

Correlation of alternate VEGF ligands with alternate angiogenic and inflammatory cytokines.

<p>Several cytokines were highly correlated with VEGF-A, VEGF-C, and PlGF. VEGF-D showed very little correlation with other cytokines. PDGF-BB was highly correlated with VEGF-A, VEGF-C, and PlGF and IFN-ɣ correlated with both VEGF-A and VEGF-C. VEGF-A and VEGF-C were also highly correlated with each other, but there was no significant correlation between the other VEGF ligands.</p

Study Schema.

<p>Study Schema.</p

Progression-free survival and Overall Survival in high and low VEGF-A and PlGF.

<p>To evaluate whether VEGF-A, VEGF-C, VEGF-D, or PlGF were prognostic, evaluable patients in the untreated group were split into two cohorts based on their median cytokine levels. VEGF-C, VEGF-D, and PlGF were not correlated with clinical outcomes. However, VEGF-A was associated with longer progression-free and overall survival. There was no correlation between the level of any VEGF ligand and the subsequent clinical outcomes in patients previously treated with or without bevacizumab.</p

DataSheet_3_Autologous humanized mouse models to study combination and single-agent immunotherapy for colorectal cancer patient-derived xenografts.xlsx

Designing studies of immunotherapy is limited due to a lack of pre-clinical models that reliably predict effective immunotherapy responses. To address this gap, we developed humanized mouse models of colorectal cancer (CRC) incorporating patient-derived xenografts (PDX) with human peripheral blood mononuclear cells (PBMC). Humanized mice with CRC PDXs were generated via engraftment of autologous (isolated from the same patients as the PDXs) or allogeneic (isolated from healthy donors) PBMCs. Human T cells were detected in mouse blood, tissues, and infiltrated the implanted PDXs. The inclusion of anti-PD-1 therapy revealed that tumor responses in autologous but not allogeneic models were more comparable to that of patients. An overall non-specific graft-vs-tumor effect occurred in allogeneic models and negatively correlated with that seen in patients. In contrast, autologous humanized mice more accurately correlated with treatment outcomes by engaging pre-existing tumor specific T-cell populations. As autologous T cells appear to be the major drivers of tumor response thus, autologous humanized mice may serve as models at predicting treatment outcomes in pre-clinical settings for therapies reliant on pre-existing tumor specific T-cell populations.</p

DataSheet_2_Autologous humanized mouse models to study combination and single-agent immunotherapy for colorectal cancer patient-derived xenografts.xlsx

Designing studies of immunotherapy is limited due to a lack of pre-clinical models that reliably predict effective immunotherapy responses. To address this gap, we developed humanized mouse models of colorectal cancer (CRC) incorporating patient-derived xenografts (PDX) with human peripheral blood mononuclear cells (PBMC). Humanized mice with CRC PDXs were generated via engraftment of autologous (isolated from the same patients as the PDXs) or allogeneic (isolated from healthy donors) PBMCs. Human T cells were detected in mouse blood, tissues, and infiltrated the implanted PDXs. The inclusion of anti-PD-1 therapy revealed that tumor responses in autologous but not allogeneic models were more comparable to that of patients. An overall non-specific graft-vs-tumor effect occurred in allogeneic models and negatively correlated with that seen in patients. In contrast, autologous humanized mice more accurately correlated with treatment outcomes by engaging pre-existing tumor specific T-cell populations. As autologous T cells appear to be the major drivers of tumor response thus, autologous humanized mice may serve as models at predicting treatment outcomes in pre-clinical settings for therapies reliant on pre-existing tumor specific T-cell populations.</p

DataSheet_1_Autologous humanized mouse models to study combination and single-agent immunotherapy for colorectal cancer patient-derived xenografts.pdf

Designing studies of immunotherapy is limited due to a lack of pre-clinical models that reliably predict effective immunotherapy responses. To address this gap, we developed humanized mouse models of colorectal cancer (CRC) incorporating patient-derived xenografts (PDX) with human peripheral blood mononuclear cells (PBMC). Humanized mice with CRC PDXs were generated via engraftment of autologous (isolated from the same patients as the PDXs) or allogeneic (isolated from healthy donors) PBMCs. Human T cells were detected in mouse blood, tissues, and infiltrated the implanted PDXs. The inclusion of anti-PD-1 therapy revealed that tumor responses in autologous but not allogeneic models were more comparable to that of patients. An overall non-specific graft-vs-tumor effect occurred in allogeneic models and negatively correlated with that seen in patients. In contrast, autologous humanized mice more accurately correlated with treatment outcomes by engaging pre-existing tumor specific T-cell populations. As autologous T cells appear to be the major drivers of tumor response thus, autologous humanized mice may serve as models at predicting treatment outcomes in pre-clinical settings for therapies reliant on pre-existing tumor specific T-cell populations.</p

Figure 2

<p>Unsupervised hierarchical clustering of ampullary adenocarcinoma samples, n = 14 (A).Clinical and molecular characteristics are listed below the figure: group 3 gene expression grouping (black), poor differentiation (black), mucinous histology (black), T4/T3 (black), N1 (black), presence of an adenoma (black), activation mutations in KRAS, BRAF, PI3K (black), MSI-high status (black), CK7+/CK20− (black), CDX-2+ (black), and histological subtype. Overall survival by gene expression derived biliary-like and intestinal-like ampullary subgroups (B).</p

Unsupervised hierarchical clustering of the differentially expressed proteins (P<0.05) between gene expression derived biliary-like and intestinal-like ampullary subgroups.

<p>Unsupervised hierarchical clustering of the differentially expressed proteins (P<0.05) between gene expression derived biliary-like and intestinal-like ampullary subgroups.</p

<i>Streptococcus gallolyticus</i> subsp. <i>gallolyticus</i> promotes colorectal tumor development

<div><p><i>Streptococcus gallolyticus</i> subsp. <i>gallolyticus</i> (<i>Sg</i>) has long been known to have a strong association with colorectal cancer (CRC). This knowledge has important clinical implications, and yet little is known about the role of <i>Sg</i> in the development of CRC. Here we demonstrate that <i>Sg</i> promotes human colon cancer cell proliferation in a manner that depends on cell context, bacterial growth phase and direct contact between bacteria and colon cancer cells. In addition, we observed increased level of β-catenin, c-Myc and PCNA in colon cancer cells following incubation with <i>Sg</i>. Knockdown or inhibition of β-catenin abolished the effect of <i>Sg</i>. Furthermore, mice administered with <i>Sg</i> had significantly more tumors, higher tumor burden and dysplasia grade, and increased cell proliferation and β-catenin staining in colonic crypts compared to mice receiving control bacteria. Finally, we showed that <i>Sg</i> is present in the majority of CRC patients and is preferentially associated with tumor compared to normal tissues obtained from CRC patients. These results taken together establish for the first time a tumor-promoting role of <i>Sg</i> that involves specific bacterial and host factors and have important clinical implications.</p></div