Last updated: 2018-08-29

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Expand here to see past versions:
    File Version Author Date Message
    Rmd dc78a95 davismcc 2018-08-29 Minor updates to analyses.
    html e573f2f davismcc 2018-08-27 Build site.
    html 9ec2a59 davismcc 2018-08-26 Build site.
    html 36acf15 davismcc 2018-08-25 Build site.
    Rmd d618fe5 davismcc 2018-08-25 Updating analyses
    html 090c1b9 davismcc 2018-08-24 Build site.
    html d2e8b31 davismcc 2018-08-19 Build site.
    html 1489d32 davismcc 2018-08-17 Add html files
    Rmd a847774 davismcc 2018-08-17 Using “line” instead of “donor”
    Rmd 1b44d28 davismcc 2018-08-13 Adding simulation analysis file.
    Rmd 1cbadbd davismcc 2018-08-10 Updating analyses.
    Rmd 2531565 davismcc 2018-08-08 Tweaking clone prevalences
    Rmd 7397e00 davismcc 2018-08-08 Updating stylez and tweaking Rmds
    Rmd 5a9a5ba davismcc 2018-08-08 Adding cowplot
    Rmd 2a45547 davismcc 2018-08-08 Adding viridis library
    Rmd d6b3b74 davismcc 2018-08-08 Adding clone prevalence analysis


Load libraries and data

knitr::opts_chunk$set(echo = TRUE)
dir.create("figures/clone_prevalences", showWarnings = FALSE, recursive = TRUE)
library(tidyverse)
library(viridis)
library(cowplot)

Load the Canopy clone inference results and the cell assignment results from cardelino for 32 donor fibroblast cell lines.

params <- list()
params$callset <- "filt_lenient.cell_coverage_sites"
fls <- list.files("data/sces")
fls <- fls[grepl(params$callset, fls)]
lines <- gsub(".*ce_([a-z]+)_.*", "\\1", fls)

cell_assign_list <- list()
for (don in lines) {
    cell_assign_list[[don]] <- readRDS(file.path("data/cell_assignment", 
        paste0("cardelino_results.", don, ".", params$callset, ".rds")))
    cat(paste("reading", don, "\n"))
}
reading euts 
reading fawm 
reading feec 
reading fikt 
reading garx 
reading gesg 
reading heja 
reading hipn 
reading ieki 
reading joxm 
reading kuco 
reading laey 
reading lexy 
reading naju 
reading nusw 
reading oaaz 
reading oilg 
reading pipw 
reading puie 
reading qayj 
reading qolg 
reading qonc 
reading rozh 
reading sehl 
reading ualf 
reading vass 
reading vils 
reading vuna 
reading wahn 
reading wetu 
reading xugn 
reading zoxy 
canopy_list <- list()

prev_list <- list()
for (don in lines) {
  tmp_df <- data_frame(
    line = don,
    clone = rownames(cell_assign_list[[don]]$tree$P),
    prev_canopy = cell_assign_list[[don]]$tree$P[, 1],
    prev_cardelino = NA,
    n_cells = length(cell_assign_list[[don]]$clone_assigned),
    n_assigned = sum(cell_assign_list[[don]]$clone_assigned != "unassigned"),
    prop_assigned = n_assigned / n_cells
  )
  for (i in seq_len(nrow(tmp_df))) {
    tmp_df$prev_cardelino[i] <- (sum(
      cell_assign_list[[don]]$clone_assigned == tmp_df$clone[i]) / 
        tmp_df$n_assigned[i])
  }
  prev_list[[don]] <- tmp_df
}
df_prev <- do.call("rbind", prev_list)

lm_eqn <- function(df) {
    m <- lm(prev_cardelino ~ prev_canopy, df);
    eq <- substitute(italic(r)^2~"="~r2, 
         list(a = format(coef(m)[1], digits = 2), 
              b = format(coef(m)[2], digits = 2), 
             r2 = format(summary(m)$r.squared, digits = 3)))
    as.character(as.expression(eq));                 
}

## Fit weighted regressions
fits <- df_prev %>%
  group_by(clone) %>% 
  do(fit = lm(prev_cardelino ~ prev_canopy, weights = prop_assigned, data = .))

fits_1grp <- df_prev %>%
  do(fit = lm(prev_cardelino ~ prev_canopy, weights = prop_assigned, data = .))

le_lin_fit <- function(dat) {
  the_fit <- lm(prev_cardelino ~ prev_canopy, weights = prop_assigned, dat)
  setNames(data.frame(t(coef(the_fit))), c("x0", "x1"))
}

fits_me <- df_prev %>%
  group_by(clone) %>% 
  do(le_lin_fit(.))

fits_me_1grp <- df_prev %>%
  do(le_lin_fit(.))

Plot clone prevalences

Plot the estimated clone fractions from the cells assigned to a clone by cardelino against the estimated clone fractions from Canopy.

fits_1grp %>%
  broom::augment(fit) %>% 
  inner_join(., df_prev) %>%
ggplot(aes(x = prev_canopy, y = prev_cardelino, shape = clone, 
                    fill = prop_assigned)) +
  geom_abline(slope = 1, intercept = 0, colour = "gray40", linetype = 2) +
  geom_ribbon(aes(ymin = .fitted - 1.645 * .se.fit, ymax = .fitted + 1.645 * .se.fit),
              fill = "gray70", alpha = 0.7) +
  geom_abline(aes(intercept = x0, slope = x1), 
              data = fits_me_1grp,
              colour = "firebrick", size = 2) +
  geom_point(size = 3) +
  xlim(0, 1) + ylim(0, 1) +
  geom_text(x = 0.9, y = 0, colour = "black", label = lm_eqn(df_prev), 
            size = 5, parse = TRUE, data = df_prev[1,]) +
  scale_fill_viridis(name = "fraction of\ncells assigned", limits = c(0, 1)) +
  scale_shape_manual(values = 21:25) +
  xlab("Estimated clone prevalence (Canopy)") +
  ylab("Assigned clone fraction (cardelino)")
Joining, by = c("prev_cardelino", "prev_canopy")

Expand here to see past versions of plot-prev-1.png:
Version Author Date
d2e8b31 davismcc 2018-08-19

ggsave("figures/clone_prevalences/clone_prev_scatter.png", 
       height = 5, width = 7)
ggsave("figures/clone_prevalences/clone_prev_scatter.pdf", 
       height = 5, width = 7)

We can also look at the same plot as above, but now faceted by the different clones.

fits %>%
  broom::augment(fit) %>% 
  inner_join(., df_prev) %>%
  ggplot(aes(x = prev_canopy, y = prev_cardelino)) +
  geom_abline(slope = 1, intercept = 0, colour = "gray40", linetype = 2) +
  geom_ribbon(aes(ymin = .fitted - 1.645 * .se.fit, ymax = .fitted + 1.645 * .se.fit),
              fill = "gray70", alpha = 0.7) +
  geom_abline(aes(intercept = x0, slope = x1), 
              data = fits_me,
              colour = "firebrick", size = 2) +
  geom_point(aes(fill = prop_assigned), size = 3, shape = 21) +
  xlim(0, 1) + ylim(0, 1) +
  facet_wrap(~clone) +
  scale_fill_viridis(name = "fraction of\ncells assigned", limits = c(0, 1)) +
  scale_shape_manual(values = 21:25) +
  xlab("Estimated clone prevalence (Canopy)") +
  ylab("Assigned clone fraction (cardelino)")
Joining, by = c("clone", "prev_cardelino", "prev_canopy")

Expand here to see past versions of plot-prev-facet-clone-1.png:
Version Author Date
d2e8b31 davismcc 2018-08-19

ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone.png", 
       height = 7, width = 9)
ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone.pdf", 
       height = 7, width = 9)

Since there are so few lines with four clones we can also make a version of the figure above with just clone1, clone2 and clone3 and fitted a weighted regression line, with points weighted by the fraction of cells assigned for the line.

fits %>%
  broom::augment(fit) %>% 
  inner_join(., df_prev) %>%
  dplyr::filter(clone != "clone4") %>%
  ggplot(aes(x = prev_canopy, y = prev_cardelino, fill = prop_assigned)) +
  geom_abline(slope = 1, intercept = 0, colour = "gray40", linetype = 2) +
  geom_ribbon(aes(ymin = .fitted - 1.645 * .se.fit, ymax = .fitted + 1.645 * .se.fit),
              fill = "gray70", alpha = 0.7) +
  geom_abline(aes(intercept = x0, slope = x1), 
              data = dplyr::filter(fits_me, clone != "clone4"),
              colour = "firebrick", size = 1) +
  geom_point(size = 3, shape = 21) +
  xlim(0, 1) + ylim(0, 1) +
  facet_wrap(~clone, nrow = 1) +
  scale_fill_viridis(name = "fraction of\ncells assigned", limits = c(0, 1)) +
  scale_shape_manual(values = 21:25) +
  xlab("Estimated clone prevalence (Canopy)") +
  ylab("Assigned clone fraction (cardelino)") +
  theme(axis.text = element_text(size = 9))
Joining, by = c("clone", "prev_cardelino", "prev_canopy")

Expand here to see past versions of plot-prev-facet-clone-3clones-1.png:
Version Author Date
d2e8b31 davismcc 2018-08-19

ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4.png", 
       height = 4.5, width = 8.5)
ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4.pdf", 
       height = 4.5, width = 8.5)

Let us also make a version of the plot above with the line joxm highlighted as this line is used as an example in the paper.

fits %>%
  broom::augment(fit) %>% 
  inner_join(., df_prev) %>%
  dplyr::filter(clone != "clone4") %>%
  dplyr::mutate(labs = ifelse(line == "joxm", "joxm", "")) %>%
  ggplot(aes(x = prev_canopy, y = prev_cardelino, fill = prop_assigned)) +
  geom_abline(slope = 1, intercept = 0, colour = "gray40", linetype = 2) +
  geom_ribbon(aes(ymin = .fitted - 1.645 * .se.fit, ymax = .fitted + 1.645 * .se.fit),
              fill = "gray70", alpha = 0.7) +
  geom_abline(aes(intercept = x0, slope = x1), 
              data = dplyr::filter(fits_me, clone != "clone4"),
              colour = "firebrick", size = 1) +
  ggrepel::geom_label_repel(aes(label = labs), fill = "gray90", size = 2.5,
                            box.padding = 0.1, label.padding = 0.15) +
  geom_point(size = 3, shape = 21) +
  xlim(0, 1) + ylim(0, 1) +
  facet_wrap(~clone, nrow = 1) +
  scale_fill_viridis(name = "fraction of\ncells assigned", limits = c(0, 1)) +
  scale_shape_manual(values = 21:25) +
  scale_colour_manual(values = c("black", "firebrick"), guide = FALSE) +
  xlab("Estimated clone prevalence (Canopy)") +
  ylab("Assigned clone fraction (cardelino)") +
  theme(axis.text = element_text(size = 9))
Joining, by = c("clone", "prev_cardelino", "prev_canopy")

ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4_joxmlabel.png", 
       height = 4.5, width = 8.5)
ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4_joxmlabel.pdf", 
       height = 4.5, width = 8.5)

ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4_joxmlabel_wide.png", 
       height = 4.5, width = 13)
ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4_joxmlabel_wide.pdf", 
       height = 4.5, width = 13)

Also look at what happens if we filter out lines that have fewer than 75% of cells assigned (25 lines).

df_prev %>%
  dplyr::filter(clone != "clone4", prop_assigned > 0.75) %>%
  ggplot(aes(x = prev_canopy, y = prev_cardelino, shape = clone, 
             fill = prop_assigned)) +
  geom_abline(slope = 1, intercept = 0, colour = "gray40", linetype = 2) +
  geom_smooth(aes(group = 1), method = "lm", colour = "firebrick") +
  geom_point(size = 3) +
  xlim(0, 1) + ylim(0, 1) +
  facet_wrap(~clone, nrow = 1) +
  scale_fill_viridis(name = "fraction of\ncells assigned", limits = c(0, 1)) +
  scale_shape_manual(values = 21:25) +
  xlab("Estimated clone prevalence (Canopy)") +
  ylab("Assigned clone fraction (cardelino)")

Expand here to see past versions of plot-prev-facet-clone-3clones-linefilt-1.png:
Version Author Date
d2e8b31 davismcc 2018-08-19

ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4_75pctassigned.png", 
       height = 4.5, width = 10.5)
ggsave("figures/clone_prevalences/clone_prev_scatter_facet_clone_no_clone4_75pctassigned.pdf", 
       height = 4.5, width = 10.5)

Session information

devtools::session_info()
Session info -------------------------------------------------------------
 setting  value                       
 version  R version 3.5.1 (2018-07-02)
 system   x86_64, darwin15.6.0        
 ui       X11                         
 language (EN)                        
 collate  en_GB.UTF-8                 
 tz       Europe/London               
 date     2018-08-29                  
Packages -----------------------------------------------------------------
 package     * version date       source        
 assertthat    0.2.0   2017-04-11 CRAN (R 3.5.0)
 backports     1.1.2   2017-12-13 CRAN (R 3.5.0)
 base        * 3.5.1   2018-07-05 local         
 bindr         0.1.1   2018-03-13 CRAN (R 3.5.0)
 bindrcpp    * 0.2.2   2018-03-29 CRAN (R 3.5.0)
 broom         0.5.0   2018-07-17 CRAN (R 3.5.0)
 cellranger    1.1.0   2016-07-27 CRAN (R 3.5.0)
 cli           1.0.0   2017-11-05 CRAN (R 3.5.0)
 colorspace    1.3-2   2016-12-14 CRAN (R 3.5.0)
 compiler      3.5.1   2018-07-05 local         
 cowplot     * 0.9.3   2018-07-15 CRAN (R 3.5.0)
 crayon        1.3.4   2017-09-16 CRAN (R 3.5.0)
 datasets    * 3.5.1   2018-07-05 local         
 devtools      1.13.6  2018-06-27 CRAN (R 3.5.0)
 digest        0.6.16  2018-08-22 CRAN (R 3.5.0)
 dplyr       * 0.7.6   2018-06-29 CRAN (R 3.5.1)
 evaluate      0.11    2018-07-17 CRAN (R 3.5.0)
 forcats     * 0.3.0   2018-02-19 CRAN (R 3.5.0)
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 git2r         0.23.0  2018-07-17 CRAN (R 3.5.0)
 glue          1.3.0   2018-07-17 CRAN (R 3.5.0)
 graphics    * 3.5.1   2018-07-05 local         
 grDevices   * 3.5.1   2018-07-05 local         
 grid          3.5.1   2018-07-05 local         
 gridExtra     2.3     2017-09-09 CRAN (R 3.5.0)
 gtable        0.2.0   2016-02-26 CRAN (R 3.5.0)
 haven         1.1.2   2018-06-27 CRAN (R 3.5.0)
 hms           0.4.2   2018-03-10 CRAN (R 3.5.0)
 htmltools     0.3.6   2017-04-28 CRAN (R 3.5.0)
 httr          1.3.1   2017-08-20 CRAN (R 3.5.0)
 jsonlite      1.5     2017-06-01 CRAN (R 3.5.0)
 knitr         1.20    2018-02-20 CRAN (R 3.5.0)
 labeling      0.3     2014-08-23 CRAN (R 3.5.0)
 lattice       0.20-35 2017-03-25 CRAN (R 3.5.1)
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 magrittr      1.5     2014-11-22 CRAN (R 3.5.0)
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 methods     * 3.5.1   2018-07-05 local         
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 munsell       0.5.0   2018-06-12 CRAN (R 3.5.0)
 nlme          3.1-137 2018-04-07 CRAN (R 3.5.1)
 pillar        1.3.0   2018-07-14 CRAN (R 3.5.0)
 pkgconfig     2.0.2   2018-08-16 CRAN (R 3.5.0)
 plyr          1.8.4   2016-06-08 CRAN (R 3.5.0)
 purrr       * 0.2.5   2018-05-29 CRAN (R 3.5.0)
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 rlang         0.2.2   2018-08-16 CRAN (R 3.5.0)
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 tidyverse   * 1.2.1   2017-11-14 CRAN (R 3.5.0)
 tools         3.5.1   2018-07-05 local         
 utils       * 3.5.1   2018-07-05 local         
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 whisker       0.3-2   2013-04-28 CRAN (R 3.5.0)
 withr         2.1.2   2018-03-15 CRAN (R 3.5.0)
 workflowr     1.1.1   2018-07-06 CRAN (R 3.5.0)
 xml2          1.2.0   2018-01-24 CRAN (R 3.5.0)
 yaml          2.2.0   2018-07-25 CRAN (R 3.5.1)

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