how to create DTW ?

[SinghalDivya]

Hi,

could you please explain how can I carry run DTW time delay analysis across modalities? as mentioned in the paper, I ran RunChromVAR but what's the input file used in example code? https://github.com/welch-lab/MultiVelo/blob/main/Examples/dtw_example.R

Thank you

[SinghalDivya]

Hi,

could you please suggest me how can I create figure 6E-G from the paper https://www.nature.com/articles/s41587-022-01476-y/figures/6.

Thanks

[danielee0707]

The input of DTW script is a file with three columns: latent time, motif accessibility, and TF gene expression. The accessibilities and expressions of genes of interest can be extracted from ATAC and RNA anndata objects respectively. The latent time is in the adata_rna.obs['latent_time'] field after computing latent time.

The paper's method section has more detailed descriptions of how Figure 6e-g were generated. For Fig6f, basically the DTW analyses were done for all TFs individually and summarized to a dataframe, then the figure was generated with ggplot like this

ggplot(data = diff_df[1:bins,], aes(x=t)) + 
  geom_line(aes(y = X0., color = 'X0.'), size=1, alpha=0.4) + 
  geom_line(aes(y = X25., color = 'X25.'), size=1.5, alpha=0.8) + 
  geom_line(aes(y = median, color = 'median'), size=2) + 
  geom_line(aes(y = X75., color = 'X75.'), size=1.5, alpha=0.8) + 
  geom_line(aes(y = X100., color = 'X100.'), size=1, alpha=0.4) + 
  geom_line(aes(y = zeros), color='black', size=1, linetype = "dashed") + 
  geom_ribbon(aes(ymin = X0., ymax = X25.), fill = '#5DB3FF', alpha=0.2) +
  geom_ribbon(aes(ymin = X25., ymax = median), fill = '#74E3AA', alpha=0.5) +
  geom_ribbon(aes(ymin = median, ymax = X75.), fill = '#C4D857', alpha=0.5) +
  geom_ribbon(aes(ymin = X75., ymax = X100.), fill = '#FF964D', alpha=0.2) +
  theme_bw() + xlab('Latent time') + ylab('Δt') +
  theme(axis.text = element_text(size=10), axis.title = element_text(size=12), legend.position = c(0.87, 0.87), 
        legend.background = element_rect(fill=NA), legend.text = element_text(size=10)) + 
  scale_color_manual("", 
                     breaks = c("X100.", "X75.", "median", "X25.", "X0."),
                     labels = c("q100", "q75", "q50", "q25", "q0"),
                     values = c("#ff5e42", "#ffcd58", "#89e255", "#5ee4ff", "#5c82ff"))

Fig6g was analyzed the same way but for SNPs. After saving all time lags to df, the plot was generated with ggplot as

ggplot(df, aes(x = snp_time, y = max_lags, color=log_count)) + geom_point(size=1, alpha=0.9) + 
  xlab('Max SNP accessibility time') + ylab('Δt') + 
  scale_color_viridis_c() + theme_bw() + theme(axis.text = element_text(size=10), axis.title = element_text(size=12))

[SinghalDivya]

Hi, Thank you so much for your response. Could you please also give me an example on how can I extract accessibilities and expressions of genes of interest can be extracted from ATAC and RNA anndata objects. Is there any sample input file for DTW analysis? Thanks again! On Aug 28, 2023, at 3:54 PM, Chen Li ***@***.***> wrote: The input of DTW is a file with three columns: latent time, motif accessibility, and TF gene expression. The accessibilities and expressions of genes of interest can be extracted from ATAC and RNA anndata objects respectively. The latent time is in the adata_rna.obs['latent_time'] field after computing latent time. The paper's method section has more detailed descriptions of how Figure 6e-g were generated. For Fig6f, basically the DTW analyses were done for all TFs and summarized to a dataframe, then the figure was generated with ggplot like this ggplot(data = diff_df[1:bins,], aes(x=t)) + geom_line(aes(y = X0., color = 'X0.'), size=1, alpha=0.4) + geom_line(aes(y = X25., color = 'X25.'), size=1.5, alpha=0.8) + geom_line(aes(y = median, color = 'median'), size=2) + geom_line(aes(y = X75., color = 'X75.'), size=1.5, alpha=0.8) + geom_line(aes(y = X100., color = 'X100.'), size=1, alpha=0.4) + geom_line(aes(y = zeros), color='black', size=1, linetype = "dashed") + geom_ribbon(aes(ymin = X0., ymax = X25.), fill = '#5DB3FF', alpha=0.2) + geom_ribbon(aes(ymin = X25., ymax = median), fill = '#74E3AA', alpha=0.5) + geom_ribbon(aes(ymin = median, ymax = X75.), fill = '#C4D857', alpha=0.5) + geom_ribbon(aes(ymin = X75., ymax = X100.), fill = '#FF964D', alpha=0.2) + theme_bw() + xlab('Latent time') + ylab('Δt') + theme(axis.text = element_text(size=10), axis.title = element_text(size=12), legend.position = c(0.87, 0.87), legend.background = element_rect(fill=NA), legend.text = element_text(size=10)) + scale_color_manual("", breaks = c("X100.", "X75.", "median", "X25.", "X0."), labels = c("q100", "q75", "q50", "q25", "q0"), values = c("#ff5e42", "#ffcd58", "#89e255", "#5ee4ff", "#5c82ff")) Fig6g was analyzed the same way but for SNPs. After saving all time lags to df, the plot was generated with ggplot with ggplot(df, aes(x = snp_time, y = max_lags, color=log_count)) + geom_point(size=1, alpha=0.9) + xlab('Max SNP accessibility time') + ylab('Δt') + scale_color_viridis_c() + theme_bw() + theme(axis.text = element_text(size=10), axis.title = element_text(size=12)) — Reply to this email directly, view it on GitHub<#32 (comment)>, or unsubscribe<https://github.com/notifications/unsubscribe-auth/AGLFXDCR4XPXYKWBZXXJNGDXXTZQJANCNFSM6AAAAAA33NY6PY>. You are receiving this because you authored the thread.Message ID: ***@***.***>

[danielee0707]

For each defined target and motif pair you can use a script similar to this to save the accessibility and expression. Depending on your specific goal (e.g. chromatin priming), the order of accessibility and expression may be flipped.

motif_activity = sc.read_csv('motifs/chromvar_z.csv')  # motif accessibility computed using chromvar
motif_activity = motif_activity.transpose()
motif_activity = motif_activity[adata_result.obs_names,:]
mv.knn_smooth_chrom(motif_activity, nn_idx, nn_dist)

for i,gene in enumerate(targets):
    t = np.reshape(np.array(adata_result.obs['latent_time']), (-1,1))
    m = np.array(motif_activity[:,motif_names[i]].layers['Mc'])
    r = np.array(adata_result[:,gene].X)
    res = np.hstack((t,m,r))
    np.savetxt('motifs/dtw/'+gene+'_res.txt', res, fmt='%f')

[SinghalDivya]

Thank you for guiding me through the process.
as suggested, I ran through the code. although I stumble upon an error:

motif_activity = sc.read_csv('./Motifname_chromvar_score_2520.csv') # motif accessibility computed using chromvar
motif_activity = motif_activity.transpose()
motif_activity = motif_activity[adata_result.obs_names,:]
mv.knn_smooth_chrom(motif_activity, nn_idx, nn_dist)

####### error #######

####### code used

motif_activity
AnnData object with n_obs × n_vars = 4060 × 746
layers: 'Mc'
obsp: 'connectivities'
####### calculated for motif (m)

calculated for r and t

###########
expression data is getting stored as sparse matrix which is 2 dimension, rest values of m and t are one dimension, if I am not wrong. could you suggest me way around this error?

Thanks again!

[danielee0707]

I think the issue here is due to adata.X being a sparse matrix. You can try r = np.array(adata_result[:,gene].X.A) instead and see if it works out.

[SinghalDivya]

Perfect, thank you it worked!