## Replication Study: Biomechanical remodeling of the microenvironment by stromal caveolin-1 favors tumor invasion and metastasis <br> **Abstract:** As part of the [Reproducibility Project: Cancer Biology][1], we published a Registered Report ([Fiering et al., 2015][2]), that described how we intended to replicate selected experiments from the paper "Biomechanical remodeling of the microenvironment by stromal caveolin-1 favors tumor invasion and metastasis" ([Goetz et al., 2011][3]). Here we report the results. Primary mouse embryonic fibroblasts (pMEFs) expressing caveolin 1 (Cav1WT) demonstrated increased extracellular matrix remodeling *in vitro* compared to Cav1 deficient (Cav1KO) pMEFs, similar to the original study ([Goetz et al., 2011][4]). *In vivo*, we found higher levels of intratumoral stroma remodeling, determined by fibronectin fiber orientation, in tumors from cancer cells co-injected with Cav1WT pMEFs compared to cancer cells only or cancer cells plus Cav1KO pMEFs, which were in the same direction as the original study (Supplemental Figure S7C; [Goetz et al., 2011][5]), but not statistically significant. Primary tumor growth was similar between conditions, like the original study (Supplemental Figure S7Ca; [Goetz et al., 2011][6]). We found metastatic burden was similar between Cav1WT and Cav1KO pMEFs, while the original study found increased metastases with Cav1WT (Figure 7C; [Goetz et al., 2011][7]); however, the duration of our *in vivo* experiments (45 days) were much shorter than in the study by [Goetz et al., 2011][8] (75 days). This makes it difficult to interpret the difference between the studies as it is possible that the cells required more time to manifest the difference between treatments observed by Goetz et al. We also found a statistically significant negative correlation of intratumoral remodeling with metastatic burden, while the original study found a statistically significant positive correlation (Figure 7Cd; [Goetz et al., 2011][9]), but again there were differences between the studies in terms of the duration of the metastasis studies and the imaging approaches that could have impacted the outcomes. Finally, we report meta-analyses for each result. ---------- ### Contents **Reports:** Read the [Replication Study][10] or view the [earlier versions][11]. **Note**: In order to successfully run and knit the Replication Study Manuscript R Markdown file, you must install a series of necessary R packages. You can review the necessary packages included in the checkpoint manifest in the markdown file [here][12] or run and knit the following r markdown file and they will be downloaded automatically. To reproduce the Replication Study manuscript text run this in [R Studio][13] (note: downloads [R markdown file][14] directly from osf.io: library(httr) library(rmarkdown) GET("https://osf.io/rd3yf/?action=download",write_disk("Replication_Study_20.Rmd", overwrite = T)) render("Replication_Study_20.Rmd", output_format = "word_document") <br> Also, explore the Registered Report and materials related to the Registered Report [here][15]. **Data and Material Availability:** All associated data, protocols, analysis scripts, and other digital materials are available within this OSF project. The image analysis workflow generated during this study (figure of workflow [here][16]) is available on Amazon Web Services (AWS) as an Amazon Machine Image (AMI). The machine image is located in the N. Virginia (us-east-1) region with the AMI ID: ami-09ee55780b0c19120, and AMI Name: rpcb-analysis-study20. Computation was performed on an Instance Type of m5.4xlarge (16 vCPU, 64 GiB Memory), with 500 GiB of Elastic Black Store (EBS) storage, and running Windows Server 2016. The administrator account password required to login is "RPCB!Analysis". **Experiments replicated**: Reproduce and explore the figures, analyses, data, and methods generated in this replication attempt. <br> - [Isolation and characterization of Cav1 WT and Cav1 KO primary MEFs][17] <br> -Replication of Figure 7Ca and Supplemental Figure S2A of Goetz et al., 2011 - [Subcutaneous tumorigenicity assay of tumor cells co-injected with Cav1 WT or Cav1 KO primary MEFs][18] <br> -Replication of Figures 7C and Supplemental Figure S7C of Goetz et al., 2011 **Meta-analysis**: As a measure of evaluating reproducibility a meta-analysis of each effect was performed. The forest plots, analyses, and data are available [here][19]. <br> Questions about the project can be directed to [contact+rpcb@cos.io.][20] [1]: https://osf.io/e81xl/wiki/home/ [2]: https://elifesciences.org/articles/04796 [3]: https://doi.org/10.1016/j.cell.2011.05.040 [4]: https://doi.org/10.1016/j.cell.2011.05.040 [5]: https://doi.org/10.1016/j.cell.2011.05.040 [6]: https://doi.org/10.1016/j.cell.2011.05.040 [7]: https://doi.org/10.1016/j.cell.2011.05.040 [8]: https://doi.org/10.1016/j.cell.2011.05.040 [9]: https://doi.org/10.1016/j.cell.2011.05.040 [10]: https://elifesciences.org/articles/45120 [11]: https://osf.io/wq32s/ [12]: https://osf.io/rd3yf/ [13]: https://www.rstudio.com [14]: https://osf.io/rd3yf/ [15]: https://osf.io/q3e4u/wiki/home/ [16]: https://osf.io/yr57n/ [17]: https://osf.io/na5h2/wiki/home/ [18]:https://osf.io/bq54u/wiki/home/ [19]:https://osf.io/rvf57/wiki/home/ [20]:mailto:contact+rpcb@cos.io